CN115612114B - Copolymer membrane and method for enzymatic self-assembly synthesis and application thereof - Google Patents

Copolymer membrane and method for enzymatic self-assembly synthesis and application thereof Download PDF

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CN115612114B
CN115612114B CN202110797790.0A CN202110797790A CN115612114B CN 115612114 B CN115612114 B CN 115612114B CN 202110797790 A CN202110797790 A CN 202110797790A CN 115612114 B CN115612114 B CN 115612114B
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CN115612114A (en
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姜文侠
田晓丽
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Tianjin Institute of Industrial Biotechnology of CAS
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Abstract

The copolymer film and the method for synthesizing the copolymer film by the enzymatic self-assembly are prepared at the interface of two liquid phases by the enzymatic reaction of a film-forming monomer containing phenolic hydroxyl, a film-forming monomer containing at least two amino groups and a catalyst. The preparation method has the advantages of simple operation steps, mild reaction conditions and wide selection range of the film forming monomers, and can select the film forming monomers according to application requirements to prepare the copolymer film with corresponding functions. The preparation method of the invention is easy to upgrade from laboratory to industrialized mass production, and has better application prospect.

Description

Copolymer membrane and method for enzymatic self-assembly synthesis and application thereof
Technical Field
The invention belongs to a novel polymer material, belongs to the fields of modern chemistry, biology, material science and application science and engineering thereof, and particularly relates to a copolymer membrane and a method and application thereof for enzymatic self-assembly synthesis at a liquid-liquid interface.
Background
Millions of organic matters are found, but the types of monomers which can be truly used as synthetic polymer raw materials are quite limited, and at most hundreds of organic matters are available, so that two or more monomers are utilized to carry out collocation copolymerization in different modes and different proportions to obtain various copolymers with different properties so as to meet different use requirements [ Wang Huai three, kou Xiaokang ]. The polymer chemistry teaching [ M ]. Beijing: scientific press, 2007:213] is still an important development direction. The design and enzymatic synthesis of novel functional polymer materials, the synthesis and application of the nano polymer are all the leading edges of polymer research and development.
The two-dimensional material and the quasi-two-dimensional material have wide application prospects in the front fields of quantum information, artificial intelligence, integrated circuits, life health, brain science, aerospace science and technology and the like, as well as the fields of sensors, logic switches, pharmaceutical preparations, medical diagnosis, electronic information, flexible wearing equipment, intelligent clothing, food safety detection, environmental protection and intelligent packaging. Films with thicknesses on the order of nanometers (1-100 nm), which may be referred to as "nanofilms", may exhibit nanomaterial properties.
The large-scale synthesis of high-quality films with wide adjustability will promote the development of artificial solids with design functions. However, many "bottom-up" polymer film fabrication techniques have significant drawbacks in terms of controllability and the like. The control of the thickness of many layers, particularly the thickness of nanofilms and their uniformity, remains a difficulty in process implementation. Uncontrollable reactions are difficult to realize in large scale commercial production and fewer processes are available to make ultra-thin multilayer films of organic polymer composites.
The stability of the film is very important for the application of the film. LB film (film of monolayer or multi-molecular layer with orderly arranged molecules prepared by Langmuir-Blodgett film forming technology) is evenly and completely regularly arranged on the molecular level. However, the mechanical strength of the LB film deposited with small molecules is not high (the LB film deposited on the substrate is scratched with a force of only several newtons), and the heat resistance, the solvent resistance and the environmental resistance are poor, and practical application is limited. In order to overcome these disadvantages, it is effective to introduce a group capable of further polymerization reaction into a film-forming small organic molecule compound to make it high-molecular [ He Pingsheng: polymerization of monolayer and Langmuir-Blodgett film [ M ]. Co-fertilizer: chinese university of science and technology Press, 2008:58.]. However, since the LB film requires amphipathic property of the film-forming organic compound monomer molecules, and the molecular chain length is limited by suitable conditions, the selection range of LB film-forming monomers is greatly limited, and difficulty is brought to the design of LB film-forming materials. Moreover, since the polymerization of monomers into polymers is a transition from van der Waals interaction distance to chemical bond distance, the shortening of the distance between molecules leads to shrinkage of the formed monolayer polymer or polymerized LB film, and the overall order of the film is not ensured [ He Pingsheng: polymerization of monolayer and Langmuir-Blodgett film [ M ]. Co-fertilizer: chinese university of science and technology Press, 2008:61.].
The supermolecular material is a modern novel material in development stage, and is based on supermolecular chemistry, and is prepared by utilizing intermolecular non-covalent bond linkage (such as hydrogen bond interaction, electron donor-acceptor interaction, ion interaction and the like) [ Zhang Zhijie ], material physical chemistry [ M ]. Beijing: chemical industry press 2020:53.]. The self-assembled membrane is an ordered supermolecule system formed by spontaneous adsorption of active molecules on heterogeneous interfaces through chemical bonds.
The development of more kinds of copolymer films and simple preparation methods thereof meets specific application requirements and becomes the direction of researchers.
Disclosure of Invention
The invention provides a method for synthesizing a copolymer film, the copolymer film prepared by the method and application thereof.
The invention relates to a method for preparing an organic copolymer film by polymerizing and self-assembling two kinds of film-forming monomers under the catalysis of a catalyst at two mutually-insoluble liquid phase interfaces (liquid-liquid interfaces), wherein one kind of film-forming monomers is an organic compound containing phenolic hydroxyl groups, and the other kind of film-forming monomers is an organic compound containing at least two amino groups. The organic copolymer film can introduce other molecules and/or atoms into the surface of the film through secondary and/or repeated chemical and/or enzymatic reactions to obtain the expected chemical composition, molecular structure, physical property and chemical property, realize the performance of the film and meet the application requirements.
In order to improve the technical problems, the invention provides a method for synthesizing a copolymer film, which comprises the following steps:
and (2) dissolving a film-forming monomer containing phenolic hydroxyl groups, a film-forming monomer containing at least two amino groups and a catalyst in an aqueous phase and/or a liquid phase which is mutually insoluble in water, and polymerizing at the interface of the two phases to form the copolymer film.
Preferably, the copolymer film is obtained by dissolving a film-forming monomer containing a phenolic hydroxyl group and a film-forming monomer containing at least two amino groups in a solvent, adding a catalyst, mixing to obtain a water phase, contacting the water phase with a liquid phase which is mutually insoluble with water, and polymerizing at the interface of the two phases to form a film.
Preferably, the film-forming monomer containing phenolic hydroxyl groups is dissolved in a solvent, a catalyst is added and mixed to obtain an aqueous phase, the film-forming monomer containing at least two amino groups is dissolved in a liquid phase which is mutually insoluble with water, the aqueous phase is contacted with the liquid phase which is mutually insoluble with water to generate enzymatic reaction, and the copolymer film is polymerized at the interface of the two phases to form the film.
Preferably, the film-forming monomer containing at least two amino groups is dissolved in a solvent, a catalyst is added and mixed to obtain an aqueous phase, the film-forming monomer containing phenolic hydroxyl groups is dissolved in a liquid phase which is mutually insoluble with water, the aqueous phase is contacted with the liquid phase which is mutually insoluble with water, and the copolymer film is polymerized at the interface of the two phases to obtain the copolymer film.
Preferably, the catalyst is dissolved in an aqueous phase, a film-forming monomer containing phenolic hydroxyl groups and a film-forming monomer containing at least two amino groups are dissolved in a liquid phase which is mutually insoluble with water, the aqueous phase is contacted with the liquid phase which is mutually insoluble with water to generate enzymatic reaction, and the copolymer film is polymerized at the interface of the two phases to form the film.
According to an embodiment of the present invention, the aqueous phase is an aqueous solution having a water content of not less than 50% by total mass; the film forming monomer can be dissolved and can contain a certain amount of organic solvent which is dissolved in water, wherein the organic solvent can be at least one of methanol, ethanol, isopropanol, acetone, methyl formate, ethyl acetate, acetonitrile, tetrahydrofuran, N-dimethylformamide, 1, 4-dioxane, dimethyl sulfoxide, diethylene glycol butyl ether and diethylene glycol;
according to an embodiment of the present invention, the solvent is selected from water or a buffer solution, and a buffer solution and buffer pair of the buffer solution are pH buffers suitable for supporting the polymerization reaction, not limited to any specific pH buffer. The buffer solution is preferably a sodium acetate-acetic acid buffer solution, disodium hydrogen phosphate-citric acid buffer solution, potassium hydrogen phthalate-sodium hydroxide buffer solution, tartaric acid-sodium tartrate buffer solution, sodium citrate-citric acid buffer solution, trisodium phosphate-phosphoric acid buffer solution, sodium malonate-malonic acid buffer solution, sodium succinate-succinic acid buffer solution, phthalic acid-hydrochloric acid buffer solution, disodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution, disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution, dipotassium hydrogen phosphate-sodium hydroxide buffer solution, tris-hydrochloric acid buffer solution, boric acid-borax buffer solution, glycine-sodium hydroxide buffer solution.
According to an embodiment of the present invention, the liquid phase that is not water-miscible may be an oil that is liquid at ordinary temperature, an oil that is solid at a set temperature, or a liquid metal. The normal temperature liquid oil may be at least one of vegetable oil, mineral oil and synthetic water insoluble liquid. The vegetable oil can be peanut oil, soybean oil, linseed oil, castor oil, rapeseed oil, corn oil, olive oil, sesame oil, cinnamon oil, essential oil, etc. The mineral oil can be petroleum (crude oil), condensate oil, gasoline, kerosene, diesel oil, lubricating oil, transformer oil, engine oil, liquid paraffin, paraffin and coal tar. The synthetic water insoluble liquid includes at least one of various silicone oils, aromatic hydrocarbons (such as benzene, toluene, xylene, dichlorotoluene, bromobenzene, etc.), alkanes (such as pentadecane, tetradecane, tridecane, dodecane, undecane, nonane, isooctane, hexane, trichloromethane, tetrachloromethane, carbon tetrachloride, carbon disulfide, etc.), cycloalkanes (such as cyclohexane, cyclopentane, cycloheptane, etc.), ethers (such as petroleum ether, butyl ether, etc.), esters (such as butyl oleate, butyl acetate, butyl palmitate, etc.), ketones (such as 2-nonanone, methyl isobutyl ketone, 3-hexanone, etc.), organic acids (such as octanoic acid, etc.). The oil is solid at a set temperature, such as at least one of paraffin, cocoa butter, coconut oil, palm oil, and animal oil. The animal oil can be derived from pig, cattle, sheep, horse, chicken, whale, insect (such as Cera flava, insect wax, etc.), etc. Such as gallium, mercury, low melting point alloys.
According to an embodiment of the invention, the catalyst may be at least one of a peroxidase and/or an oxidoreductase having laccase activity and/or an artificial enzyme having the aforementioned catalytic activity;
according to an embodiment of the present invention, the peroxidase may be selected from at least one of manganese peroxidase (Manganese peroxidase, mnP, EC 1.11.1.13), lignin peroxidase (lin peroxidase, liP, EC 1.11.1.14) and Chloroperoxidase (Chloroperoxidase, CPO, EC 1.11.1.10), plant peroxidase (Plant peroxidase); the plant peroxidase may be horseradish peroxidase (Horseradish peroxidase, HRP, EC 1.11.1.7), soybean peroxidase (Soybean peroxidase, SBP, EC 1.11.1.7), rice peroxidase (Rice peroxidase), cotton peroxidase (Cotton peroxidase), safflower bean peroxidase (Runner beans peroxidase), chickpea peroxidase (Garbanzo beans peroxidase), guar peroxidase (Guar beans peroxidase), pea peroxidase (Pea peroxidase), and the like.
According to an embodiment of the invention, the oxidoreductase having Laccase activity is at least one of a enzyme classification EC 1.10.3.2 specified by the nomenclature committee of the international union of biochemistry and molecular biology (Nomenclature Committee of the International Union of Biochemistry and molecular Biology, IUBMB) comprising any Laccase (lacccase) from plants, animals, fungi, bacteria, yeasts or obtained by biotechnology, and/or any fragment exhibiting Laccase activity from its source, and/or an enzyme exhibiting similar activity, such as Catechol oxidase (Catechol oxidase, CO, EC 1.10.3.1) or any fragment exhibiting Catechol oxidase activity from its source, monophenol monooxidase (Monophenol monooxygenase, EC 1.14.18.1) or any fragment exhibiting monophenol monooxidase activity from its source, bilirubin oxidase (Bilirubin oxidase, BOD, EC 1.3.3.5) or any fragment exhibiting bilirubin oxidase activity from its source.
According to an embodiment of the invention, the laccase of plant origin is selected from laccase in the group consisting of Anacardiaceae (Anacardiaceae), momordica (podocarpae), aesculus (aeacculus sp.), acer palmatum (Acer pseudoplatanus), vinca (Catharanthus roseus), carrot (Daucus carrata), dichomitius squalens), ginkgo (ginko biloba), apple (Malus pumila), cymbidium (Monotropa hypopithys), avocado (Persea americana), peach (Prunus persica), potato (Solanum tuberosum), rosemary (Rosmarinus officinalis), vinca minor (Vinca minor) extracts.
According to an embodiment of the invention, the laccase is preferably a fungal laccase. The fungal laccase is selected from the group consisting of agaricus bisporus (Agaricus bisporus), aspergillus nidulans (Agaricus bisporus), botrytis cinerea (Botrytis cinerea), ceralopecuroides (Agaricus bisporus), acremonium monocolor (Cerrena unicolor), coriolus thermophilus (Agaricus bisporus), acremonium bud (Agaricus bisporus), coprinus cinereus (Agaricus bisporus), coriolus versicolor (Agaricus bisporus), phellinus linteus (Agaricus bisporus), ganoderma lucidum (Agaricus bisporus), callicarpa sativa (Agaricus bisporus), monascus pinnatifida (Agaricus bisporus), rumex crispus (Agaricus bisporus), pleurotus saxifraga (Agaricus bisporus), thermomyces lanuginosus (Agaricus bisporus), myces lanuginosus (Agaricus bisporus), phanerochaete chrysosporium (Agaricus bisporus), pleurotus roseus, pachytridactylotheca (Agaricus bisporus), pleurotus grifolius (Agaricus bisporus), hymenochaete pensis (Agaricus bisporus), hymenochaetes rupulus (Agaricus bisporus), hymenochaete, russella (Agaricus bisporus) and (Agaricus bisporus), trametes versicolor (Tramates versicolor) and laccase variants thereof.
Preferably, the fungus laccase is Polyporus winter (Polyporus brumalis) (the preservation number of the strain is CCTCC NO: M2020809) laccase.
Laccase may also be an enzyme produced by a method comprising the steps of: culturing a host cell transformed with a recombinant DNA vector carrying a DNA sequence encoding the function of the laccase in a culture medium under conditions such that the laccase is expressed, and recovering the laccase from the culture.
Fungal laccase has the following advantages over other enzymes: (1) The catalytic substrate is wide, and the enhancer added with the enzyme can catalyze more substrates; (2) a high redox potential compared to bacterial laccase; (3) the industrialized preparation is easier to realize; (4) lower cost; (5) no coenzyme is required; (6) The oxidant uses molecular oxygen, so that hydrogen peroxide is not required to be added in the reaction; (7) Water is the only byproduct and does not form an enzyme-substrate complex that is useless; (8) glycosylation modification is carried out, so that the stability of the enzyme is good; (9) low sensitivity to metal ions in the medium; (10) resistance to a number of organic solvents.
According to an embodiment of the present invention, the phenolic hydroxyl group-containing compound has a structure as shown in formula I:
wherein each R 1 The same or different are independently selected from H, halogen, CN, NO 2 OH, SH, COOH, unsubstituted or substituted by one, two or more R a1 Substituted with the following groups: c (C) 1-40 Alkyl, C 2-40 Alkenyl, C 2-40 Alkynyl, C 3-40 Cycloalkyl, C 3-40 Cycloalkenyl, C 3-40 Cycloalkynyl radicals, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, -OR 1-2 、-SR 1-3 、-NR 1-4 R 1-5 、-C(O)R 1-6 、-OC(O)R 1-7 、-S(O) 2 R 1-8 、-OS(O) 2 R 1-9 、-P(O)R 1-10 R 1-11 、-N＝NR 1-12 ；
A 1 Presence or absence; when A is 1 When present, is selected from unsubstituted or substituted by one, two or more R b1 Substituted C linked to the benzoring 6-20 Aryl, 5-20 membered heteroaryl, 5-20 membered heterocyclyl; or A 1 Selected from chemical bonds, unsubstituted or optionally substituted by one, two or more R c1 Substituted O, C (O), C (O) O, S, S (O) 2 、N、C 1-6 Alkylene, ch= N, N = N, CH =n-n=ch, ch=ch-CO-CH 2 -CO-CH＝CH；
m is an integer of 0 to 5.
Each R a1 、R b1 、R c1 Identical or different, independently of one another, from the group consisting of H, halogen, CN, OH, SH, oxo (= O), NO 2 、COOH、-OR 1-2 、-SR 1-3 、-NR 1-4 R 1-5 、-C(O)R 1-6 、-OC(O)R 1-7 、-S(O) 2 R 1-8 、-OS(O) 2 R 1-9 、P(O)R 1- 10 R 1-11 Unsubstituted or optionally substituted by one, two or more R 1-12 Substituted C 1-40 Alkyl, C 2-40 Alkenyl, C 2-40 Alkynyl, C 3-40 Cycloalkyl, C 3-40 Cycloalkenyl, C 3-40 Cycloalkynyl radicals, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl;
each R is 1-2 、R 1-3 、R 1-4 、R 1-5 、R 1-6 、R 1-7 、R 1-8 、R 1-9 、R 1-10 、R 1-11 、R 1-12 Identical or different, independently of one another, from the group consisting of H, halogen, CN, OH, SH, oxo (= O), NO 2 、COOH、C 1-40 Alkyl, C 2-40 Alkenyl, C 2-40 Alkynyl, C 3-40 Cycloalkyl, C 3-40 Cycloalkenyl, C 3-40 Cycloalkynyl radicals, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl.
According to an embodiment of the present invention, formula I has the structure shown in formulas I-1 to I-13:
wherein R is 1 M and R c1 Having the definition set forth above; b is selected from O or R 1 Substituted N; D. d (D) 1 、D 2 And D 3 Identical or different, independently of one another, from chemical bonds, unsubstituted or optionally substituted by one, two or more R c1 Substituted O, CH 2 C (O) or N-CO-CH 3 The method comprises the steps of carrying out a first treatment on the surface of the E and E 1 Identical or different, independently of one anotherSelected from chemical bonds, unsubstituted or optionally substituted by one, two or more R c1 Substituted O, C (O), C (O) O, S, S (O) 2 、N、Se、P、C 1-6 Alkylene, ch=ch, ch= N, N = N, CH =n-n=ch, ch=ch-CO-CH 2 -CO-ch=ch; n is an integer of 0 to 5; f (F) 1 And F 2 The same or different, independently of one another, are selected from N, CH or O + ；R 1 ’、R 1 "and R 1 Is the same as defined in the following.
Preferably, the compound of formula I-1 is selected from catechol (CAS: 120-80-9), 3-methylcatechol (CAS: 488-17-5), 3, 4-dihydroxytoluene (CAS: 452-86-8), 4-ethylcatechol (CAS: 1124-39-6), 4-tert-butyl-1, 2-benzenediol (CAS: 98-29-3), 3, 5-di-tert-butyl-1, 2-benzenediol (CAS: 1020-31-1), 3-methoxycatechol (CAS: 934-00-9), 3, 4-dihydroxybenzaldehyde (CAS: 139-85-5), 3, 4-dihydroxystyrene (CAS: 6053-02-7), 2',4' -dihydroxyacetophenone (CAS: 89-84-9), hydroxytyrosol (CAS: 10597-60-1), dopamine (CAS: 51-61-6) and salts thereof, 3-methacryl dopamine (CAS: 471915-89-6), epinephrine (CAS: 51-43), isopropanolamine (CAS: 139-5), 3, 4-dihydroxyacetophenone (CAS: 35-9), 5-bromocatechol (CAS: 35-35), and salts thereof, and the like, and the reagent (3, 4-dihydroxycatechol, 3-dihydroxybenzaldehyde (CAS: 39-5) may be selected from the group consisting of catechol, 3-dihydroxyacetophenone (CAS: 39-9) Esters such as ethyl 3, 4-dihydroxybenzoate (CAS: 3943-89-3), esters such as ethyl 2, 3-dihydroxybenzoate, 4-nitrocatechol (CAS: 3316-09-4), urushiol, suilin (CAS: 103538-03-0), grifolin (CAS: 6903-07-7), ilicifolin B (CAS: 22581-07-3), resorcinol (CAS: 108-46-3), 4-chlororesorcinol (CAS: 95-88-5), 2, 6-dihydroxytoluene (CAS: 608-25-3), 2, 4-dihydroxybenzaldehyde (CAS: 1995-1-2), 2',4' -dihydroxyacetophenone (CAS: 89-84-9), 2',6' -dihydroxyacetophenone (CAS: 699-83-2), 4-hexylresorcinol (CAS: 136-77-6), cardiotonic phenol (Cardol), terbutaline sulfate (CAS: 31-32-5), hydroquinone (CAS: 123-9), hydroquinone (CAS: 108-46-3), 4-chlororesorcinol (CAS: 95-88-5), 2,6 '-dihydroxyacetophenone (CAS: 35-35), 2', 35-dihydroxyacetophenone (CAS: 35-35), and 15235-6-5) Calcium dobesilate (CAS: 20123-80-2), cannabigerol (CAS: 25654-31-3), oxacinne sulfate (CAS: 5874-97-5), cannabidiol (CAS: 13956-29-1), phloroglucinol (CAS: 6099-90-7), 2,4, 6-trihydroxybenzaldehyde (CAS: 487-70-7), ethyl 2,4, 6-trihydroxybenzoate (CAS: 90536-74-6) and the like, hydrochloric acid-6-hydroxydopamine (CAS: 28094-15-7), hydrobromic acid 6-hydroxydopamine (CAS: 636-00-0), 2,4, 5-trihydroxybenzylalanine (CAS: 23358-64-7), pyrogallol (CAS: 87-66-1), 2',3',4' -trihydroxy acetophenone (CAS: 528-21-2), 2,3, 4-trihydroxybenzaldehyde (CAS: 2144-08-3), methyl 2,4, 6-trihydroxybenzoate (CAS: 99-24-1), bromoethyl bromide (CAS: 90536-74-6), and the like), methyl phenol (CAS: 106-6-5-hydroxydopamine (CAS: 35-6-hydroxy-6), methyl phenol (CAS: 106-6-5), methyl phenol (CAS: 106-6-5-hydroxy phenol) and the like, 6-hydroxy phenol (CAS: 106-6-5-methyl phenol) and the like, 2, 3-dimethoxyphenol (CAS: 5150-42-5), 4-ethylphenol (CAS: 123-07-9), vanillin (CAS: 121-33-5), isovanillin (CAS: 621-59-0), vanillyl alcohol (CAS: 498-00-0), guaiacol (CAS: 90-05-1), o-acetaminophen (CAS: 614-80-2), acetaminophen (CAS: 103-90-2), 3,4, 5-trimethoxyphenol (CAS: 642-71-7), 4- (2' -thiazolylazo) resorcinol (CAS: 2246-46-0), esters such as ethyl 2,3, 4-trihydroxybenzoate, benserazide hydrochloride (CAS: 14919-77-8), and the like.
Preferably, the compound of I-2 is selected from the group consisting of 3', 4-dihydroxyflavone (CAS: 4143-64-0), 6, 7-dihydroxyflavone (CAS: 38183-04-9), 7, 8-dihydroxyflavone (CAS: 38183-03-8), farnesin (CAS: 480-44-4), coryza (CAS: 480-40-0), kaempferin (CAS: 491-54-3), lycopene (CAS: 520-12-7), isorhamnetin (CAS: 480-19-3), geransin (CAS: 520-34-3), coumartin (CAS: 4423-37-4), daidzein (CAS: 491-71-4), valin (CAS: 480-15-9), azascine (CAS: 529-51-1), tamaxanthin (CAS: 603-61-2), 5, 7-dihydroxy-2- (4-hydroxyphenyl) -6, 8-dimethoxy-4H-1-benzopyran-4-one (CAS: 520-35-80), coumarone (CAS: 4323-37-4), azascine (CAS: 491-48), azascine (CAS: 548-35-48), homogin (CAS: 52-35-8), homogin (CAS: 52-35-8) Dihydroquercetin (CAS: 6151-25-3), 6-methoxygambogin (CAS: 520-11-6), 3-O-methyl quercetin (CAS: 1486-70-0), morin (CAS: 480-16-0), luteolin (CAS: 491-70-3), quercetin (CAS: 117-39-5), myricetin (CAS: 529-44-2), genkwanin (CAS: 437-64-9), 5, 7-dihydroxy-6,8,4 ' -trimethoxyflavone (CAS: 10176-66-6), pimannoxin (CAS: 56003-01-1), kaempferol (CAS: 520-18-3), scutellarin (CAS: 529-53-3), 7,3',4' -trihydroxy-3, 8-dimethoxyflavone, 3,7,3',4' -tetrahydroxy-8-methoxyflavone, 3,7,8,3',4' -pentahydroxyflavone, and the like.
Preferably, the compound of I-3 is selected from Dekkoxaden (CAS: 897-46-1), daidzein (CAS: 486-66-8), biochanin A (CAS: 491-80-5), 4',6, 7-trihydroxyisoflavone (CAS: 17817-31-1), genistein (CAS: 446-72-0), olol (CAS: 480-23-9), karaine (CAS: 20575-57-9), prunetin (CAS: 552-59-0), and the like.
Preferably, the compound of I-4 is selected from escin (CAS: 305-01-1), 4-methyl escin (CAS: 529-84-0), and the like.
Preferably, the compound of I-5 is selected from the group consisting of hesperetin (CAS: 520-33-2), naringenin (CAS: 67604-48-2), flavodoxin (CAS: 20725-03-5), naringenin (CAS: 480-41-1), epicatechin (CAS: 24808-04-6), robinetin (CAS: 4382-33-6), dihydroquercetin (CAS: 480-18-2), dihydromyricetin (CAS: 27200-12-0), eriodictyol (CAS: 552-58-9), non-sirolimus (CAS: 490-49-3), (+) -Rhobatanol (CAS: 17445-90-8), gallocatechin (CAS: 3371-27-5), 4', 7-dihydroxyisoflavan (CAS: 531-95-3), avermectin (CAS: 2545-00-8), (+) -catechin (CAS: 154-23-4), epigallocatechin (CAS: 970-74-1), epicatechin, broussonetianol, phenazocine, monomeric procyanidins, monomeric faberidine, monomeric procyanidins, dihydromyricetin, 3' -hydroxy-35-84, etc.
Preferably, the compound of I-6 is selected from the group consisting of anthocyanin (CAS: 528-58-5), delphinidin chloride (CAS: 528-53-0), pelargonidin (CAS: 134-04-3), cassidine chloride (CAS: 23130-31-6), petuniin (CAS: 1429-30-7), malvidin (CAS: 643-84-5), and the like.
Preferably, the compound of I-7 is selected from the group consisting of p-phenylphenol (CAS: 92-69-3), azo violet (CAS: 74-39-5), 2, 4-dihydroxybenzophenone (CAS: 131-56-6), phenethylresorcinol (CAS: 85-27-8), xanthohumol (CAS: 6754-58-1), resveratrol (CAS: 501-36-0), rosmarinic acid (CAS: 20283-92-5), phloretin (CAS: 60-82-2), nordihydroguaiaretic acid (CAS: 500-38-9), tetrahydroxy stilbene, piceatannol (CAS: 10083-24-6), curcumin (CAS: 458-37-7), bisphenol A (CAS: 80-05-7), 2 '-dihydroxybiphenyl (CAS: 1806-29-7), 4' -dihydroxybiphenyl (CAS: 92-88-6), phenolphthaline (CAS: 81-90-3), thiobischlorophenol (CAS: 97-18-7), bischlorophenol (CAS: 97-23-4), 2 '-dihydroxybenzophenone (CAS: 835-11-0), 4' -dihydroxybenzophenone (CAS: 80-35-607) and 4-99-96-5 2,2' -dihydroxy-4-methoxybenzophenone (CAS: 131-53-3), honokiol (CAS: 35354-74-6), magnolol (CAS: 528-43-8), diethylstilbestrol (CAS: 5635-50-7), diethylstilbestrol (CAS: 6898-97-1), 3', 5' - (tetraisopropyl) biphenyl 4,4' -diol (CAS: 2416-95-7), 2' -dihydroxydiphenyl ether (CAS: 15764-52-0), dobutamine hydrochloride (CAS: 49745-95-1), witch hazel tannin (CAS: 469-32-9), 2, 3', 4', 5-hexahydroxydiphenylsulfone, 3', 4', 5' -hexahydroxydiphenylsulfone, 2', 3', 4' -hexahydroxydiphenylsulfone, hexahydroxydiphenylacid, and the like.
Preferably, the compound of I-8 is selected from 2, 3-dihydroxynaphthalene (CAS: 92-44-4), 1, 2-dihydroxynaphthalene (CAS: 574-00-5), 1, 3-dihydroxynaphthalene (CAS: 132-86-5), 1, 5-dihydroxynaphthalene (CAS: 83-56-7), 1, 6-dihydroxynaphthalene (CAS: 575-44-0), 1, 7-dihydroxynaphthalene (CAS: 575-38-2), 1, 4-dihydroxynaphthalene (CAS: 571-60-8), 2, 7-dihydroxynaphthalene (CAS: 582-17-2), 6, 7-dihydroxy-2-sodium naphthalene sulfonate (CAS: 135-53-5), 3, 5-dihydroxy-2-naphthoic acid (CAS: 89-35-0), calcium carboxylic acid (CAS: 3737-95-9), calcium magnesium reagent (CAS: 3147-14-6), edestin blue R (CAS: 2538-85-4), chromium-2R (CAS: 4197-07-3), dicarboxyphenyl acid (CAS: 130-85-8), beryllium II (51550-25-5), chromic acid (CAS: 582-17-2), 6, 7-dihydroxy-2-naphthoic acid (CAS: 89-35-25), chromium (3838-35-6), chromium (3848), chromium (387-35-9), and chromium (387-35-6), chromium (387-35-9), and chromium (387-35-9) And Everest blue SE (CAS: 1058-92-0), etc.
Preferably, the compound of I-9 is selected from shikonin (CAS: 517-89-5) and the like.
Preferably, the compound of I-10 is selected from the group consisting of aloesin (CAS: 481-72-1), dithranol (CAS: 1143-38-0), 1, 5-dihydroxyanthraquinone (CAS: 117-12-4), 1, 4-dihydroxyanthraquinone (CAS: 81-64-1), 1, 8-dihydroxyanthraquinone (CAS: 117-10-2), 10-acetyl-3, 7-dihydroxyphenazine (CAS: 119171-73-2), anthralin (CAS: 117-12-4), chrysophanol (CAS: 481-74-3), rhein (CAS: 478-43-3), 1,2, 3-trihydroxyanthraquinone (CAS: 602-64-2), rhodoxanthin (CAS: 81-54-9), alizarin red (CAS: 130-22-3), alizarin complexing indicator (CAS: 3952-78-1), alizarin (CAS: 72-48-0), alizarin red S (CAS: 130-22-3), tolanthraquinone hydrochloride (70476-82-3), fluocinolone (CAS: 518-5), emodin (CAS: 518-82), and alizarin (CAS: 35-82), and alizarin (35-8-35-81-82) Alpha-mangostin (CAS: 6147-11-1), and the like.
Preferably, the compound of I-11 is selected from the group consisting of galloblue (CAS: 1562-85-2), azulene (CAS: 1562-90-9), and the like.
Preferably, the compound of I-12 is selected from 1,8, 9-trihydroxyanthracene (CAS: 480-22-8), and the like.
Preferably, the compound of I-13 is selected from the group consisting of parahydroxybenzoic acid (CAS: 99-96-7), m-hydroxybenzoic acid (CAS 99-06-9), vanillic acid (CAS 121-34-6), isovanillic acid (CAS 645-08-9), 4-hydroxy-2-methoxybenzoic acid (CAS 90111-34-5), 3-hydroxy-5-methoxybenzoic acid (CAS 19520-75-6), salicylic acid (CAS 69-72-7), p-aminosalicylic acid (CAS 65-49-6), m-aminosalicylic acid (CAS 89-57-6), 3-methylsalicylic acid (CAS 83-40-9), 5-methylsalicylic acid (CAS 89-56-5), 3-methoxysalicylic acid (CAS 877-22-5), 4-methoxysalicylic acid (CAS 2237-36-7), 5-methoxysalicylic acid (CAS 2612-02-4), 6-methoxysalicylic acid (CAS 3147-64-6), 3, 5-diiodosalicylic acid (CAS 133-91-5), syringic acid (CAS 530-57-6), eugenol (CAS 35-37-60), and 35-37-60-45-35-60, such as ginkgolic acid Diflunisal (CAS: 22494-42-4), gentisic acid (CAS: 490-79-9), pyrocatechol (CAS: 303-38-8), resorcinol acid (CAS: 89-86-1), 3, 5-dihydroxybenzoic acid (CAS: 99-10-5), 2, 6-dihydroxybenzoic acid (CAS: 303-07-1), gallic acid (CAS: 149-91-7), ferulic acid (CAS: 1135-24-6), p-coumaric acid (CAS: 501-98-4), tyrosine, sinapic acid (CAS: 530-59-6), 4-hydroxymandelic acid (CAS: 1198-84-1), 2-hydroxyphenylglycine (CAS: 25178-38-5), caffeic acid (CAS: 331-39-5), diphenolic acid (CAS: 126-00-1), protocatechuic acid (CAS: 99-50-3), danshensu (CAS: 76822-21-4), dopa, methyldopa (CAS: 555-30-6), 2,4, 6-trihydroxybenzoic acid (CAS: 5783-30-7), and the like.
According to an embodiment of the present invention, formula I also includes substances in which two or more structural units represented by formulas I-1 to I-13 are present in the structure, such as pinosylvin (CAS: 521-34-6), cinnamon tannins (CAS: 86631-38-1), 2 '-dihydroxy-1, 1' -binaphthyl (CAS: 602-09-5), 6 '-dithiodinaphthol (CAS: 6088-51-3), hyperin (CAS: 548-04-9), thiopyrimidine pamoate (CAS: 22204-24-6), 2, 3-dihydroginkgetin (CAS: 34421-19-7), amentoflavone (CAS: 1617-53-4), isoginkgetin (CAS: 548-19-6), norginkgetin (CAS: 521-32-4), 5' -methoxyginkgetin (CAS: 77053-35-1), ginkgetin (CAS: 481-46-9), procyanidin B1 (CAS: 20315-25-7), procyanidin B2 (CAS: 29106-49-8), procyanidin B3 (CAS: 23567-23-9), procyanidin B4 (CAS: 06-51-2), procyanidin B5 (CAS: 12798-57-1), procyanidin B6 (CAS: 12798-58-2), procyanidin B (CAS: 3878-59), procyanidin B8 (CAS: 12798-60-6), (-) -epigallocatechin gallate (CAS: 989-51-5), epicatechin gallate (CAS: 1257-08-5), catechin gallate (CAS: 130405-40-2), gallocatechin gallate (CAS: 4233-96-9), procyanidin B2-3' -O-gallate, dehydrocatechin B, tetrabrominated tannin, procyanidin B-2,3' -O-gallate, procyanidin B-2,3' -di-O-gallate, procyanidin B1 (CAS: 78362-04-6), procyanidin B2 (CAS: 87392-61-8), procyanidin B3 (CAS: 78362-05-7), procyanidin B4 (CAS: 68964-95-4), and the like.
According to an embodiment of the invention, the compounds of formula I further comprise: saxifragin (CAS: 477-90-7), hanbain (CAS: 632-85-9), tectorigenin (CAS: 548-77-6), licoflavone (CAS: 59870-68-7), ellagic acid (CAS: 476-66-4), catalin (CAS: 82-08-6), yunnan J (CAS: 99217-67-1), hematoxylin oxide (CAS: 475-25-2), rhodol (CAS: 569-77-7), gnetin A (CAS: 82084-87-5), pyrogallol red (CAS: 32638-88-3), hematoxylin (CAS: 517-28-2), catechol violet (CAS: 115-41-3), 1-naphtholphthalein (CAS: 596-01-0), azoarsin I (CAS: 520-10-5), azoarsin III (CAS: 1668-00-4), phenolphthalein (CAS: 77-09-8), phenol red (CAS: 74-8), tetraiodoform (CAS: 569-77-7), ganin A (CAS: 82084-87-5), pyrogallol red (CAS: 32638-88-3), hematoxylin (CAS: 517-5-125-3), hematoxylin (CAS: 596-5), 4-5-40-5), and luciferin (CAS: 596-5) Rosolic acid (CAS: 633-00-1), thymol blue (CAS 76-61-9), alizarin violet (CAS 2103-64-2), glabridin (CAS 60008-03-9), ke Pusu, anthocyanin, cuacodine, heimiolA, balanicarpol α -H, dehydrocatechin A, fulvic acid (CAS 479-66-3) and salts thereof, mycophenolic acid (CAS 24280-93-1), alizarin blue S (CAS 66675-89-6), apomorphine hydrochloride (CAS 41372-20-7), (4E) -6- (4, 6-dihydroxy-7-methyl-3-oxo-1, 3-dihydroisobenzofuran-5-yl) -4-methyl-4-hexenoic acid 2- (morpholin-4-yl) ethyl ester (CAS 1322681-36-6), (+) -boldine (CAS 70-0), pseudoclip 24-6), tubuloside (CAS 57-94-3), silybin (888-7-methyl-3), fuzosin (13292-92-1), fuzosin (13292-92-35-6), and rifampicin (13292-35-80-1) Rifamycin SV, rifaximin (CAS: 80621-81-4), doxorubicin hydrochloride (CAS: 25316-40-9), epirubicin hydrochloride (CAS: 56360-09-1), rifapentine (CAS: 61379-65-5), doxorubicin (CAS: 54193-28-1), li Fumi Tet (CAS: 2750-76-7), vancomycin and salts thereof, norvancomycin hydrochloride (CAS: 213997-73-0), yellow-3-en-3-ol, teicoplanin (CAS: 61036-62-2), malibatol A, malibatol B (CAS: 204644-72-4), suffruticoside A, suffruticoside B, vactanol C, and the like.
According to an embodiment of the invention, the compounds of formula I further comprise: polyphenols.
According to an embodiment of the present invention, the polyphenol comprises: procyanidin (CAS: 4852-22-6), procyanidin A1 (CAS: 103883-03-0), procyanidin A2 (CAS: 41743-41-3), procyanidin A4 (CAS: 111466-29-6), procyanidin C1 (CAS: 37064-30-5), procyanidin C2 (CAS: 37064-31-6), cinnamon tannin B1 (CAS: 88082-60-4), cinnamon tannin B2 (CAS: 88038-12-4), theaflavin (CAS: 4670-05-7), tea polyphenols (CAS: 84650-60-2), tannins (CAS: 1401-55-4), vain (CAS: 79786-01-9), cerin (CAS: 81739-27-7), daviciin T1 (CAS: 137371-86-9), granatin (CAS: 65995-64-4), gemin A (CAS: 137371-86), sanguisorba officinalis H2 (CAS: 4630-52-35), tea polyphenols (CAS: 84650-60-35-6), tannins (CAS: 81739-35-7), davidian (CAS: 8179), davidian, and (CAS: 8234-35) Equisetum (CAS: 79786-00-8), alpha-viniferin (CAS: 62218-13-7), epsilon-viniferin (CAS: 62218-08-0), delta-viniferin, gnetin C (CAS: 84870-54-2), gnetin D (CAS: 84870-53-1), gnetin J (CAS: 152511-23-4), pallidol (CAS: 105037-88-5), uterol C (CAS: 109605-83-6), salvianolic acid B (CAS: 115939-25-8), humic acid (CAS: 1415-93-6) and salts thereof, nitro humic acid (Nitro humicic acid), huangfen, dehydrodittanoic acid, valonec acid and esters thereof, biflavanethecine, oolong tea, granin, delphinidin, and the like.
According to an embodiment of the invention, the compounds of formula I further comprise: compounds of the formula I-1 to I-13 in combination with quinic acid and/or glycosyl and/or sugar acid form, such as arbutin (CAS: 497-76-7), scutellarin (CAS: 27740-01-8), native daidzin (CAS: 155-58-8), chlorogenic acid (CAS: 327-97-9), colchicoside (CAS: 604-80-8), puerarin (CAS: 3681-99-0), hesperidin (CAS: 520-26-3), neohesperidin (CAS: 13241-33-3), diosmin (CAS: 520-27-4), aloin (CAS: 5133-19-7), forsythoside E (CAS: 93675-88-8), angoroside C (CAS: 115909-22-3), kaempferol-7-O-D-glucoside (CAS: 16290-07-6), naringin (CAS: 36-47-2), apigenin-7-glucoside (CAS: 13241-33-3), diosmin (CAS: 520-27-4), aloin (CAS: 5133-19-7), forsythin E (CAS: 93675-88-8-9), angustol C (CAS: 115909-22-3), kaempferol-7-O-D-glucoside (CAS: 16290-37-9), amycin (CAS: 35-39-4), and/or 8-10 kaempferol-3-O-glucose (1-2) rhamnoside (CAS: 142451-65-8), flubenine (CAS: 55804-74-5), isorhamnetin-3-O-D-glucoside (CAS: 5041-82-7), syringin-3-O-D-rutinoside (CAS: 53430-50-5), luteolin-4 '-glucoside (CAS: 6920-38-3), astilbin (CAS: 29838-67-3), chloroglucoside paeoniflorin (CAS: 6906-39-4), colchicine-3-O-galactoside (CAS: 30113-37-2), malvidin-3-O-arabinoside (CAS: 28500-04-1), phlorizin (CAS: 60-81-1), neohesperidin dihydrochalcone (CAS: 20702-77-6), rutin (CAS: 153-18-4), trobin II (CAS: 81571-4), catechin-7, 3' -O-beta-D-glucoside, bis-glucopyranoside-7-D-glucopyranoside-7-giucomatoside-7-D-arabinoside-7-giucomatoside, and bis-O-glucopyranoside-7-D-glucopyranoside-7-giucomatoside, quercetin-4-methoxy-3 '-D-glucoside, catechin-4' -O-beta-D-glucose, triemarin II, agrimonine, geraniin phenazine, tea gallol, shrimp flower glucoside, ma Sangyin, poly-flavanoloxy glucoside, poly-flavanolcarboside, quercetin-3-O-beta-D-glucose, 3-O- {2-O- [6-O- (p-D-glucosyl) -O-anti-coumaroyl ] -D-glucosyl } - (L-rhamnosyl) quercetin, rosacin A, rosacin B, rosacin C, rosacin D, rosacin E, rosacin F, rosacin G, luteolin 7-O-rhamnose glucoside, luteolin 7-O-glucose glucoside, genistein 7-O-rhamnose glucoside, quercetin-3-O-glucoside, lobal 7-O-rhamnose glucoside, 3-O- [2-O, 6-O-oxaziram-3-O-glucose, quercetin-3-O-2-O-glucose, oxabisporin-3 '-O-2' -O-glucopyranosyl) -2 '-D-glucopyranosyl-glucoside, quercetin-2' -O-alpha-2 '-O-glucopyranosyl-4' -glucopyranosyl-p-glucopyranoside, and the like. The glycosyl includes glucose, xylose, galactose, arabinose, rhamnose, mannose, glucuronic acid, rutinose, gentiobiose, sophorose, neohesperidose, locust disaccharide, lactose, sophorose, caffeoyl glucose, 2-acetylglucose, and combinations thereof.
Preferably, the film-forming monomer containing phenolic hydroxyl groups is selected from at least one of 1, 3-dihydroxynaphthalene, p-hydroxybenzoic acid, ferulic acid, 1, 6-dihydroxynaphthalene, shikonin, 3-methylsalicylic acid, p-coumaric acid, 4 '-dihydroxybiphenyl, bisphenol a, chlorogenic acid, 3-methoxysalicylic acid, gallic acid, 4',6, 7-trihydroxyisoflavone, geraniin, tannic acid, 3, 5-diiodosalicylic acid, gentisic acid, resveratrol, esculentin, p-aminosalicylic acid, caffeic acid, ellagic acid, pinbiflavone, anthocyanin, syringic acid, tyrosine, guaiacol, 2, 6-dihydroxytoluene, catechol, hydroxytyrosol, hesperetin, 2,4, 6-trihydroxybenzaldehyde, chrysin, tectorigenin, colchicoside, rhein, galloblue, 1,8, 9-trihydroxyanthracene, urushiol, tert-butylhydroquinone.
According to an embodiment of the present invention, the film-forming monomer containing at least two amino groups has a structure represented by formula II:
each R 2 The same or different are independently selected from H, halogen, CN, NO 2 NO, OH, SH, COOH, unsubstituted or substituted by one, two or more R a2 Substituted with the following groups: c (C) 1-40 Alkyl, C 2-40 Alkenyl, C 2-40 Alkynyl, C 3-40 Cycloalkyl, C 3-40 Cycloalkenyl, C 3-40 Cycloalkynyl radicals, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, -OR 2-2 、-SR 2-3 、-NR 2-4 R 2-5 、-C(O)R 2-6 、-OC(O)R 2-7 、-S(O) 2 R 2-8 、-OS(O) 2 R 2-9 、P(O)R 2-10 R 2-11 ；
A 2 Selected from unsubstituted or substituted by one, two or more R b2 Substituted C linked to the benzoring 1-40 Alkyl, C 2-40 Alkenyl, C 2-40 Alkynyl, C 3-40 Cycloalkyl, C 3-40 Cycloalkenyl, C 3-40 Cycloalkynyl radicals, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl; or A 2 Selected from chemical bonds, unsubstituted or optionally substituted by one, two or more R c2 Substituted O, C (O), C (O) O, S, S (O) 2 、N、C 1-6 Alkylene, ch= N, N = N, CH =n-n=ch, ch=ch-CO-CH 2 -CO-CH＝CH；
p is an integer of 0 to 12;
each R a2 、R b2 、R c2 Identical or different, independently of one another, from the group consisting of H, halogen, CN, OH, SH, oxo (=o), =nh, NO 2 、COOH、C 1-40 Alkyl, C 2-40 Alkenyl, C 2-40 Alkynyl, C 3-40 Cycloalkyl, C 3-40 Cycloalkenyl, C 3-40 Cycloalkynyl radicals, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, -OR 2-2 、-SR 2-3 、-NR 2-4 R 2-5 、-C(O)R 2-6 、-OC(O)R 2-7 、-S(O) 2 R 2-8 、-OS(O) 2 R 2-9 、P(O)R 2-10 R 2-11 ；
Each R is 2-2 、R 2-3 、R 2-4 、R 2-5 、R 2-6 、R 2-7 、R 2-8 、R 2-9 、R 2-10 、R 2-11 The same or different, independently of one another, from H, halogen, NH 2 CN, OH, SH, oxo (=o), NO 2 、COOH、C 1-40 Alkyl, C 2-40 Alkenyl, C 2-40 Alkynyl, C 3-40 Cycloalkyl, C 3-40 Cycloalkenyl, C 3-40 Cycloalkynyl radicals, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl.
According to an embodiment of the present invention, formula II has the structure shown in formulas II-1 to II-10:
Wherein R is 2 P and R c2 Having the definition set forth above; q is an integer of 0 to 12; r is R 2 ’、R 2 "and R 2 Is the same as defined in the specification; and R is 2 、R 2 ’、R 2 "has at least two amino groups therein;
each D 2 、D 3 、D 4 Identical or different, independently of one another, from N, unsubstituted or substituted by R 2 Substituted CH, N + ；
Each E 2 、E 3 、E 4 Identical or different, independently of one another, from chemical bonds, unsubstituted or optionally substituted by one, two or more R c2 Substituted O, C (O), C (S), C (O) O, S, S (O) 2 、N、C 1-6 Alkylene, ch= N, N = N, CH =n-n=ch, C 0-6 alkylene/alkenyl-CO-C 1-6 alkylene-CO-C 0-6 Alkylene/alkenyl, C 0-6 alkylene/alkenyl-CO-NH-C 0-6 alkylene-CO-C 0-6 Alkylene/alkenyl, C 0-6 alkylene/alkenyl-NH-C 0-6 alkylene-NH-C 0-6 Alkylene/alkenyl, C 0-6 alkylene/alkenyl-C (O) O-C 0-6 alkylene-C (O) O-C 0-6 Alkylene/alkenyl groups.
Each F 2 The same or different, independently of one another, are selected from O, S.
Preferably, the compound of II-1 is selected from the group consisting of p-phenylenediamine (CAS: 106-50-3) and salts thereof, p-toluenediamine (CAS: 95-70-5), 2, 3-Dimethyl-p-phenylenediamine (2, 3-Dimethyl-1, 4-benzonediamine), 2, 6-Dimethyl-p-phenylenediamine (2, 6-Dimethyl-1, 4-benzonediamine), 2, 3-Diethyl-p-phenylenediamine (2, 3-Dimethyl-1, 4-benzonediamine), 2, 5-Dimethyl-p-phenylenediamine (CAS: 6393-01-7), 2-hydroxyethyl-p-phenylenediamine sulfate (CAS: 93841-25-9), 2-Fluoro-p-phenylenediamine (2-Fluoro-1, 4-benzonediamine), 2-Isopropyl-p-phenylenediamine (2-Isopropyl-1, 4-benzonediamine), 2-Hydroxymethyl-p-phenylenediamine (2-hydroymethyl-1, 4-benzonediamine), 2-Hydroxyethoxy-p-phenylenediamine (2-hydroyethoxy-1, 4-benzonediamine), 2-acetamido-p-phenylenediamine (2-acetamidoxy-1, 4-benzodiazinediamine), N- (4-aminobenzyl) biguanide, 2-nitro-1, 4-phenylenediamine (CAS: 5-dichloro-1, 4-phenylenediamine (CAS: 615-09), 2-dichloro-1, 4-phenylenediamine (CAS: 615-09-7), 2, 5-toluenediamine sulfate (CAS: 615-50-9), o-phenylenediamine (CAS: 95-54-5) and salts thereof, 4-chloro-1, 2-phenylenediamine (CAS: 95-83-0), amiloride hydrochloride (CAS: 2016-88-8), 4, 5-dichloro-1, 2-phenylenediamine (CAS: 5348-5), 3, 4-diaminoanisole (CAS: 496-72-0), 3-nitro-1, 2-phenylenediamine (CAS: 3694-52-8), 4-nitro-1, 2-phenylenediamine (CAS: 99-56-9), 3, 4-diaminobenzoic acid (CAS: 619-05-6), meta-phenylenediamine (CAS: 108-45-2) and salts thereof, 4-chloro-1, 3-phenylenediamine (CAS: 5131-60-2), 2, 6-diaminotoluene (CAS: 823-40-5), 2, 4-diaminoanisole (CAS: 615-05-0), 3-nitro-1, 2-phenylenediamine (CAS: 3694-52-8), 4-nitro-1, 2-diaminobenzoic acid (CAS: 99-56-9), 3, 4-diaminobenzoic acid (CAS: 619-05-6), 3, 5-diaminobenzoic acid (CAS: 108-45-2), 3-diaminobenzoic acid (CAS: 35-35), 3, 35-diaminobenzene, 3-2-diaminobenzene (CAS: 35-35), and salts thereof 2, 4-diamino-6-methyl-1, 3, 5-triazine (CAS: 542-02-9), melamine (CAS: 108-78-1), 6-phenyl-1, 3, 5-triazine-2, 4-diamine (CAS: 91-76-9), veratrine (CAS: 5355-16-8), 2,4, 5-triamino-6-hydroxypyrimidine sulfate (CAS: 35011-47-3), trimethoprim (CAS: 738-70-5), phenazopyridine hydrochloride (CAS: 136-40-3), pyrimethamine (CAS: 58-14-0), eostiradine maleate (CAS: 84504-69-8), trimethoprim (CAS: 738-70-5), 4-amino-6-chlorobenzene-1, 3-disulfonamide (CAS: 121-30-2), metanilide, and the like.
Preferably, the compound of II-2 is selected from 3, 4-diaminobenzophenone (CAS: 39070-63-8), basic orange 2 (CAS: 532-82-1), o-tolidine (CAS: 119-93-7) and salts thereof, 3 '-diaminobenzidine (CAS: 91-95-2) and salts thereof, 4' -diaminobiphenyl (CAS: 92-87-5) and salts thereof, 3', 5' -tetramethylbenzidine dihydrochloride (CAS: 54827-17-7), bis (4-amino-3-chlorophenyl) methane (CAS: 101-14-4), 4 '-diaminodiphenylmethane (CAS: 101-77-9), 4' -diaminodiphenyl ether (CAS: 101-80-4), 4 '-diaminodiphenyl sulfide (CAS: 139-65-1), 4' -diaminobenzophenone (CAS: 611-98-3), 4 '-diaminodibenzyl (CAS: 621-95-4), 4' -diaminodiphenyl sulfone (CAS: 80-08-0), 4 '-diaminodiphenylamine and salts thereof, 3-dichlorobenzidine (CAS: 91-94-1), 3' -dimethoxybenzidine (CAS: 101-80-4), 4 '-diaminodiphenyl sulfide (CAS: 139-65-1), 4' -diaminobenzidine (CAS: 611-98-3), 4 '-diaminobenzidine (CAS: 621-95-4), 4' -diaminobenzidine (CAS: 80-0) 4,5' -diaminodiphenyl sulfide, and the like.
Preferably, the compound of II-3 is selected from ethylenediamine (CAS: 107-15-3) and salts thereof, 1, 2-propanediamine (CAS: 78-90-0), 1, 3-propanediamine (CAS: 109-76-2), 1, 4-diaminobutane (CAS: 110-60-1), 1, 5-pentanediamine (CAS: 462-94-2), 1, 6-hexanediamine (CAS: 124-09-4), 1, 7-heptanediamine (CAS: 646-19-5), 1, 8-octanediamine (CAS: 373-44-4), 1, 10-decanediamine (CAS: 646-25-3), diethylenetriamine (CAS: 111-40-0), triethylenetetramine (CAS: 112-24-3), tetraethylenepentamine (CAS: 112-57-2), pentaethylenehexamine (CAS: 4067-16-7), hexamine, spermidine (CAS: 124-20-9), homospermidine (CAS: 4427-76-3), aminopropyl cadaverine (CAS: 56-19-9), norspermine (CAS: 56-18-8), spermine (CAS: 71-44-3), thermophilic spermine (CAS: 70862-2), thermophilic spermine (CAS: 13274-35-379), homophilic pentamine (CAS) Thermophilic hexylamine (CAS: 63833-74-9), thermophilic homohexylamine (CAS: 133416-04-3), 3 '-iminodipropylamine (CAS: 56-18-8), N' -bis (3-aminopropyl) -1, 3-propanediamine (CAS: 4605-14-5), urea (CAS: 57-13-6), thiosemicarbazide, biuret (CAS: 108-19-0), thiosemicarbazide (CAS: 79-19-6), acrylamide methylene urea, guanidine (CAS: 90332-86-8), biguanide nitrate (CAS: 22817-07-8), glycylglutamine (CAS: 13115-71-4), alanylglutamine (CAS: 39537-23-0), amide, N- (2-guanidine ethyl) guanidine (CAS: 44956-51-6), 2, 6-diaminopimelic acid (CAS: 583-93-7), 2, 4-diaminobutyric acid dihydrochloride, annine (CAS: 57-53-4), 2, 3-diaminopropionic acid (CAS: 58-94-35-6), amino guanidine hydrochloride, amino guanidine (Ammonine-35-60), amino guanidine hydrochloride, and the like.
Preferably, the compound of II-4 is selected from the group consisting of procyanidin (CAS: 92-62-6) and salts thereof, safranine T (CAS: 477-73-6), and the like.
Preferably, the compound of II-5 is selected from 6-hydroxy-2, 4, 5-triaminopyrimidine (CAS: 1004-75-7), 2, 4-diamino-6-hydroxypyrimidine (CAS: 1956-6-4), and the like.
Preferably, the compound of II-6 is selected from 5, 6-diamino-1, 3-dimethyluracil (CAS: 5440-00-6), 4, 5-diamino-6-hydroxy-2-mercaptopyrimidine (CAS: 1004-76-8), and the like.
Preferably, the compound of II-7 is selected from the group consisting of 2, 3-diaminonaphthalene (CAS: 771-97-1), 1, 8-diaminonaphthalene (CAS: 479-27-6), 1, 5-diaminonaphthalene (CAS: 2243-62-1), triamterene (CAS: 396-01-0), methotrexate (CAS: 59-05-2), dihydralazine sulfate (CAS: 7327-87-9), and the like.
Preferably, the compound of II-8 is selected from 1, 2-diaminocyclohexane (CAS: 694-83-7), streptavidin (CAS: 488-52-8), and the like.
Preferably, the compound of II-9 is selected from 4,4' -diaminodicyclohexylmethane (CAS: 1761-71-3), and the like.
Preferably, the compound of II-10 is selected from the group consisting of 1, 2-diaminoanthraquinone (CAS: 1758-68-5), 1, 5-diaminoanthraquinone (CAS: 129-44-2), 1, 4-diaminoanthraquinone (CAS: 128-95-0), and the like.
According to an embodiment of the present invention, the compound of formula II also includes substances in which two or more structural units represented by formulas II-1 to II-10 are present in the structure, such as 4,4 '-binaphthyl amine (CAS: 481-91-4), 3' -dimethylbinaphthyl amine (CAS: 13138-48-2), and the like.
According to an embodiment of the invention, the compounds of formula II also include polymers having a plurality of amino groups in the structure, such as: polyvinyl amine (CAS: 49553-92-6), polyethylenimine (CAS: 9002-98-6), and the like.
According to an embodiment of the invention, the compound of formula II further comprises: direct brown 44 (CAS: 6252-62-6), direct blue 1 (CAS: 2610-05-1), mecobalamin (CAS: 13422-55-4), vitamin B 12 (CAS: 68-19-9), ethidium bromide (CAS: 1239-45-8), 3, 5-diamino-1, 2, 4-triazole (CAS: 1455-77-2), neored (CAS: 3248-91-7), parared base (CAS: 467-62-9), parared and salts thereof, amitriptyline (CAS: 37691-11-5), asparagine, lysine and salts thereof, hydroxylysine, cystine and salts thereof, arginine and salts thereof, ornithine and salts thereof, selenocysteine and salts thereof, 2, 7-diaminofluorene (CAS: 525-64-4), congo red (CAS: 573-58-0), valacyclovir hydrochloride (CAS: 136489-37-7), thyme pentapeptide (CAS: 69558-55-0), 4 '(5') -diaminodibenzo-15-crown-5 (CAS: 245086-08-2), adenosylcobalamine (CAS: 70-90-1), (Z) -2- (2-aminothiazol-4-yl) -2- (hydroxyimino) acetamide (CAS: 1450758-21-0), polymyxin B (CAS: 136489-37-7), thymosin pentapeptide (CAS: 69558-55-0), 4 '(5') -diaminodibenzoxacin hydrochloride (CAS: 1404-28-35-37-8), amicin hydrochloride (CAS: 37-35-8), and salts thereof) Streptomycin sulfate (CAS: 3810-74-0), neomycin sulfate (CAS: 1405-10-3), bleomycin (CAS: 11056-06-7), kanamycin, mitomycin (CAS: 50-07-7), sisomicin sulfate (CAS: 53179-09-2), gentamycin sulfate, isopalmitin sulfate (CAS: 67814-76-0), colicin sulfate (CAS: 1405-37-4), nisin (CAS: 3930-19-6), 5, 6-diamino-2, 4-dihydroxypyrimidine sulfate dihydrate (CAS: 63981-35-1), famotidine (Famotidine), 4', 6-diamidino-2-phenylindole, and the like.
Preferably, the film forming monomer containing at least two amino groups is selected from the group consisting of arginine, polyethylenimine (PEI), kanamycin, 2-fluoro-p-phenylenediamine, 2, 3-dimethyl-p-phenylenediamine, diethylenetriamine, lysine, 3-nitro-1, 2-phenylenediamine, p-phenylenediamine, 2, 3-diaminonaphthalene, procyanidin, 2, 3-diaminopyridine, 2, 3-diethyl-p-phenylenediamine, 2, 4-diaminobenzenesulfonic acid, paramaranthrene, 1, 6-hexamethylenediamine, ethidium bromide, 2-hydroxymethyl-p-phenylenediamine, 2-hydroxyethoxy-p-phenylenediamine, 5, 6-diamino-1, 3-dimethyluracil, melamine, m-phenylenediamine, 3, 4-diaminotoluene, spermine, 4' -diaminodibenzyl, 3, 4-diaminobenzoic acid, 2, 4-diaminoanisole, o-tolidine, 4' -diaminodicyclohexylmethane, 4' -dinaphthylamine, urea, guanidine, 6-hydroxy-2, 4, 5-diaminopyrimidine, 1, 2-diaminocyclohexane, 2, 4-diamino-triazine, 1, 4-diamino-4-diphenyl sulfone, 4-diamino-1, 4-diamino-sulfone.
According to an embodiment of the present invention, when the catalyst is at least one of laccase, bilirubin oxidase and peroxidase, the combination of the film-forming monomer containing a phenolic hydroxyl group and the film-forming monomer containing at least two amino groups may be: combining: r is R 1 And/or A 1 A combination of a compound of formula I and a compound of formula II when there is at least one phenolic hydroxyl group in the structure; or two in combination: compounds of formula I-13 and when D 2 Is R is 2 Substituted CH, and R 2 、R 2 ’、R 2 "a combination of compounds of formula II-1 when there are at least two amino groups in the compound.
According to an embodiment of the present invention, when the catalyst is a monophenol monooxidase, the combination of the film-forming monomer containing a phenolic hydroxyl group and the film-forming monomer containing at least two amino groups may be: a combination of a compound of formula I and a compound of formula II.
According to an embodiment of the present invention, when the catalyst is catechol oxidase, the combination of the film-forming monomer containing a phenolic hydroxyl group and the film-forming monomer containing at least two amino groups may be: r is R 1 A combination of a compound of formula I and a compound of formula II when the compound is a phenolic hydroxyl group in an ortho position.
According to an embodiment of the present invention, the film-forming monomer containing a phenolic hydroxyl group and the film-forming monomer containing at least two amino groups may be one, two or more, respectively, in the reaction system.
According to an embodiment of the present invention, the enzymatic reaction may further incorporate at least one of an enhancer for enzymes, hydrogen peroxide, metal ions, buffer ion pairs;
Preferably, the method comprises the steps of, the enzyme enhancer comprises alpha-Raffinic acid, beta-Raffinic acid, gamma-Raffinic acid, cinnamic acid, 4-hydroxy cinnamic acid, needle alcohol, ethyl vanillin, caffeic acid, ferulic acid, 2, 4-pentanedione, 4' -dihydroxybenzophenone, benzoic acid, sodium benzoate, 3-hydroxybenzoic acid, 4-hydroxybenzoic acid, 3-hydroxy anthranilic acid, 3-hydroxy-2-aminobenzoic acid, 4-amino-3-hydroxybenzoic acid, 2, 3-hydroxybenzoic acid, 3, 4-dihydroxybenzoic acid, dimethoxybenzoic acid, 2,4, 6-trihydroxybenzoic acid, p-hydroxyphenylacetic acid, salicylic acid, salicylate, 4-amino-salicylic acid, pyruvic acid, pyruvate, nicotinic acid, ascorbic acid ascorbate, guanidine, cyanuric acid, imidazole, 2, 6-dimethylphenol, 2,4, 6-trimethylphenol, 2-acetaminophen, p-coumaric acid, 7-hydroxycoumarin, sinapic acid, syringaldehyde, syringic acid, methyl syringate, ethyl syringate, propyl syringate, butyl syringate, hexyl syringate, octyl syringate, vanillic acid, isovanillic acid, vanillin even nitrogen, acetosyringone, acetovanillone, vanillyl alcohol, homovanillic acid, catechol, epicatechin, naringin, tyrosine, 2-thiouracil, ethyl 3- (4-hydroxy-3, 5-dimethoxyphenyl) acrylate, quercetin, 2,2' -Azino-bis- (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (2, 2' -Azino-bis (3-ethylbenzidine-6-sulfonic acid) diammonium salt, ABTS), hydroxydiphenylethanol, dimethoxybenzyl alcohol, N-hydroxyphthalimide, N-acetyl-N-phenylhydroxylamine, N-benzylidene-benzylamine, N-hydroxy-N-acetanilide, N-hydroxybutyrylanilide, 4' -dimethoxy-N-methyl-diphenylamine, 4' -diaminodiphenylamine, triphenylamine, benzidine, N-benzylidene-4-benzidine, 3' -dimethylbenzidine, 3' -dimethoxybenzidine, 3',5,5' -tetramethylbenzidine, 4' -dihydroxybiphenyl, 4' -hydroxy-4-biphenylcarboxylic acid, phenol red, 2', 6' -tetramethylpiperidine oxide, 1- (3 ' -sulfophenyl) -3-methyl-5-pyrazolone, purpuric acid, p-hydroxybenzosulfonate, 1, 5-diaminonaphthalene, 7-methoxy-2-naphthol, 6-hydroxy-2-naphthoic acid, 7-amino-2-naphthalenesulfonic acid, 5-amino-2-naphthalenesulfonic acid, 7-hydroxy-1, 2-naphthoimidazole, 1-hydroxybenzotriazole, 10-methylphenoxazine, 10- (2-hydroxyethyl) phenoxazine, 10-phenoxazine propionic acid, phenothiazine-10-propionic acid, 10- (2-pyrrolidinylethyl) phenothiazine, 10-allylphenothiazine, 10- (3-hydroxypropyl) phenothiazine, 2-acetyl-10-methylphenothiazine, 10-ethyl-4-phenothiazinecarboxylic acid, 10-ethylphenothiazine, 10-propylphenothiazine, 10-isopropylphenothiazine, methyl 10-phenothiazinpropionate, 10-methylphenothiazine, 10-phenothiazine-propionic acid, thiophenazine-10-propionic acid (phenothiazine-10-propiinicacid), PPT), N-hydroxysuccinimide-10-phenothiazine-propionic acid, 10- (3- (4-methyl-1-piperazinyl) propylphenothiazine, promazine, chlorpromazine, 3-hydroxy-1, 2, 3-benzotriazin-4 (3H) -one, N- (4-cyanophenyl) acetohydroxamic acid, iminostilbene, 4-amino-4 ' -methoxystilbene, 4' -diaminostilbene-2, 2' -disulfonic acid, 2, 7-diaminofluorene, 2- (p-aminophenyl) -6-methylbenzothiazole-7-sulfonic acid, N- (4- (dimethylamino) benzylidene) -p-anisaldehyde, 3-methyl-2-benzothiazolinone (4- (dimethylamino) benzylidene) hydrazone, N-hydroxyacetylene lactone, black liquor from hardwood, black liquor from softwood, wood organic solvents (ligno-organosolv), lignosulfonates, and mixtures thereof.
According to an embodiment of the present invention, the catalyst is used in the reaction system in an amount of 0.01U/L to 600U/L, for example, 0.5U/L to 200U/L, and exemplified by 1U/L, 2U/L, 6U/L, 12U/L, 24U/L, 40U/L, 60U/L, in terms of enzyme activity.
According to an embodiment of the present invention, the film-forming monomer may have a mass concentration in the reaction system of 0.002 to 380g/L, for example, 0.1 to 20g/L, and exemplified by 0.5g/L, 0.8g/L, 1g/L, 2g/L, 3g/L, 4g/L.
According to embodiments of the present invention, the film forming temperature may be 4 to 90 ℃, for example 10 to 60 ℃, illustratively 20 ℃, 30 ℃, 35 ℃, 37 ℃, 40 ℃, 45 ℃, 50 ℃, 55 ℃;
according to embodiments of the present invention, the film forming time may be 0.01 to 72 hours, for example, 2 to 48 hours, and exemplary are 3 hours, 5 hours, 6 hours, 8 hours, 10 hours, 12 hours, 20 hours, 24 hours, 36 hours;
according to embodiments of the invention, the pH of the aqueous phase may be 2 to 10, for example 3, 4, 5, 6, 6.5, 7, 8.
According to an embodiment of the invention, when the catalyst is laccase, bilirubin oxidase, monophenol monooxidase or catechol oxidase, the reaction requires oxygen to participate. The source of oxygen can be pure oxygen or mixed gas containing oxygen, further, the oxygen can be oxygen from air, can be oxygen dissolved in the surface of a solution, and can be slowly dissolved in a reaction system by flowing, mechanically stirring, oscillating, a blower and an air compressor to provide oxygen for the reaction. An oxygen-enriched liquid may be added to the liquid.
For reactions requiring hydrogen peroxide, the source of hydrogen peroxide in solution may be at least one of the following: (1) adding a hydrogen peroxide solution to the solution; (2) Adding to the solution an enzyme system that generates hydrogen peroxide, such as glucose oxidase and glucose, the glucose oxidase oxidizing glucose to generate hydrogen peroxide; galactose oxidase and galactose, galactose oxidase oxidizes galactose to produce hydrogen peroxide; sugar alcohol oxidase and glycerol, the sugar alcohol oxidase oxidizing glycerol to hydrogen peroxide; amino acid oxidase, flavin Adenine (FAD) and amino acid, the amino acid oxidase oxidizes amino acid with FAD as a prosthetic group to produce hydrogen peroxide; l-glutamic acid oxidase and L-glutamic acid, the L-glutamic acid oxidase oxidizing L-glutamic acid to produce hydrogen peroxide; (3) Hydrogen peroxide sources, i.e., compounds that produce hydrogen peroxide when dissolved in water or a suitable water-based medium, including, but not limited to, metal peroxides, t-butyl hydroperoxide, cumene hydroperoxide, percarbonates, persulfates, perphosphates, peracetic acid, perborates, perbenzoic acid, peroxyacids, alkyl peroxides, acyl peroxides, peroxyesters, urea peroxides, and peroxycarboxylic acids or salts thereof, optionally supplemented with catalase.
The interface of the two phases can be an oil-lower water phase (reaction solvent) interface of which the upper normal temperature is liquid, an upper water phase (reaction solvent) -oil (such as carbon tetrachloride, carbon disulfide and butyl palmitate) interface of which the lower normal temperature is liquid, an oil (such as paraffin, beeswax, shellac, spermaceti, cocoa butter, coconut oil, beef tallow, mutton tallow and the like) interface of which the lower temperature is solid at a set temperature, an upper water phase (reaction solvent) -lower liquid metal (such as gallium, mercury and low-melting-point alloy) interface, an interface between liquid drops of the water phase (reaction solvent) suspended in the oil and the oil, and an interface between oil drops suspended in the water phase (reaction solvent) and the water phase.
The invention also provides copolymer particles or copolymer films prepared by the synthesis method.
According to an embodiment of the invention, the copolymer film comprises the following types: the phases of the catalyst and the film-forming monomer can be classified into the following three types: (1) A film formed by dissolving the catalyst and the film-forming monomer in the water phase; (2) A catalyst and at least one film-forming monomer are dissolved in the aqueous phase, and another film-forming monomer or monomers form a film in the oil phase; (3) The catalyst is dissolved in the aqueous phase and the film-forming monomer is dissolved in the oil phase to form a film. The interface of the film can be classified into the following: a film of an interface of an upper normal temperature being liquid oil-lower water phase, a film of an interface of an upper water phase-lower normal temperature being liquid oil, a film of an interface of an upper water phase-lower liquid metal, a film of an interface of an aqueous phase-oil (said oil phase being solid at a set temperature such as paraffin, beeswax, shellac, spermaceti, cocoa butter, coconut oil, beef tallow, etc.), an oil-water interface film of a water-in-oil emulsion, a water-oil interface film of an oil-in-water emulsion.
The invention also provides applications of the copolymer particles or copolymer films, such as applications in preparing multilayer films, preparing vesicles, preparing conductive films and constructing supermolecule functional materials; and in reducing metals, coating applications; applications in secondary reactions, such as further modification of copolymer films;
preferably, the modification incorporates sorbitol, quaternary ammonium salt, dye molecules on the surface of the copolymer film.
The copolymer membranes of the present invention also include vesicles. Vesicles (Vesicles) of the invention are a class of assemblies with a closed structure surrounded by a copolymer membrane.
The invention also provides a preparation method of the vesicle, which comprises the following steps: (1) And (3) dissolving the catalyst and the film-forming monomer in the water phase, and adding the water phase and the film-forming monomer into the oil-in-water emulsion to obtain the vesicle with the surface of the copolymer film. (2) And (3) dissolving the catalyst and the film-forming monomer in a water phase, pouring the water phase and the film-forming monomer into oil, stirring to obtain water-in-oil emulsion, and reacting for a period of time to obtain the vesicle with the surface being the copolymer. (3) Dissolving a catalyst and a water-soluble film-forming monomer in a water phase, dissolving another oil-soluble monomer in oil, preparing emulsion, and reacting for a period of time to obtain the vesicle wall which is the copolymer film.
The selection of the film-forming monomer has a remarkable influence on the characteristics of the chemical composition, molecular structure, physicochemical properties, performance and the like of the copolymer film, so that the chemical composition, molecular structure, physicochemical properties, performance and the like of the copolymer film can be designed on a molecular level by selecting a proper film-forming monomer and the proportion thereof. Such as: the general organic polymer is a good insulator, and suitable film-forming monomers such as hydroquinone, ferulic acid, catechol, p-phenylenediamine, benzidine and the like with conjugated pi bond are selected, so that the copolymer film has pi electrons with high delocalization in the structure, can be used for preparing films with conductive performance, and can be used for organic high polymer full-color plane display materials and devices, optical limiting materials, sensing materials of sensors, electrode materials and microwave absorbing materials.
The present invention also provides a multilayer film comprising at least one layer of the copolymer film.
According to an embodiment of the invention, the preparation method can also be used for increasing the thickness of the membrane by supplementing the raw materials of the enzymatic reaction and/or replacing a fresh enzymatic reaction system.
Those skilled in the art will recognize that the reaction rate, film thickness, and film surface reactive functional groups can be easily controlled by changing the kind and addition amount of the enzyme, the kind and concentration of the film-forming monomer, pH, temperature, the kind and concentration of the ion pair in the buffer, the metal ion and its concentration, the concentration of the oxidizing agent (e.g., dissolved oxygen, hydrogen peroxide), the duration of the enzymatic reaction, the enhancer of the enzyme, the inhibitor of the enzymatic activity, whether the organic solvent and the kind and concentration of the organic solvent are contained, the composition of the liquid medium constituting the interface, whether the reaction solution is updated, whether and how the reaction is terminated, and the like. The thickness of the film, the thickness of the deposit/bond on the film, can be controlled individually on the nanometer scale.
In accordance with the principles of the present invention, any buffer, pH, ion, and any combination thereof may be used for the reaction system that can satisfy the copolymerization reaction. Other aspects of the enzymatic reaction conditions of the present invention can be optimized by routine experimentation. For example, pH and temperature are non-limiting examples of optimizable factors.
By selecting suitable film-forming monomers, it is possible to design and prepare copolymer films that expose specific reactive functional groups at the molecular level, i.e. residual reactive and affinity groups on the copolymer film obtained after polymerization of the monomers, which groups may be: halogen, unsubstituted or substituted by halogen, OH, COOH, NH 2 、CN、SH、NO 2 Alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl substituted with: OH, COOH, NH 2 SH, alkyl, alkoxy, alkenyl, alkynyl, aryl, sulfonyl, sulfonamide, acyl halide, phosphoryl, and azo. The active groups on the copolymer film can introduce functional molecules and/or atoms and/or ions into the surface of the copolymer film through secondary or repeated reactions, so that the further functionalization of the surface of the copolymer film is realized, the surface of the copolymer film has extremely strong designability, and the application range of the film is further expanded.
According to embodiments of the present invention, the copolymer film can be designed and prepared to expose specific reactive functional groups at the molecular level by varying the type and concentration of film-forming monomers, and the functional groups on the film can be chemically or enzymatically reacted. The modification and/or modification of the membrane is achieved by chemical or enzymatic reactions, such as radical reactions, grafting reactions, to attach the desired target molecules/atoms to the membrane. Such group reactions include, but are not limited to: michael-type addition reaction, schiff base reaction (Schiff base reaction), friedel-Crafts reaction, mannich reaction, substitution reaction, addition reaction, polymerization reaction, condensation reaction, oxidation reaction, reduction reaction, esterification reaction, sulfonation reaction, nitration reaction, diazotization reaction, azide reaction, amidation reaction, acylation reaction, alkylation reaction, glycosidation reaction, ether formation reaction, etherification reaction, coupling reaction, complexation reaction, displacement reaction, nucleophilic elimination reaction, surface atom transfer radical polymerization (SI-ATRP) reaction, reversible addition-fragmentation chain transfer polymerization (RAFT) reaction.
Such as: the proper monomer is selected so that the prepared copolymer film contains rich phenolic hydroxyl groups. Under certain conditions, the phenolic hydroxyl group is oxidized into a quinoid structure, so that the amino group can be combined with the thiol (-SH) and amino (-NH) 2 ) The hydrophilic or hydrophobic organic molecules of the polymer film undergo Michael addition and Schiff base reaction, and functional molecules are introduced into the surface of the polymer film, so that Self-assembled monolayer modification (Self-assembled monolayers) of the surface of the polymer film can be realized.
Such as: the phenolic hydroxyl groups in the copolymer film are capable of forming coordination bonds with the metal ions in the metal oxide. Immersing the copolymer film in ammonium hexafluorotitanate ((NH) 4 ) 2 TiF 6 ) And boric acid (H) 3 BO 3 ) Mixed solution of (NH) 4 ) 2 TiF 6 And H 3 BO 3 Titanium dioxide (TiO) produced by hydrolysis 2 ) Chelating the nano particles with the copolymer film to form uniform TiO on the surface of the copolymer film 2 A film.
Those skilled in the art will appreciate that: the surface of the copolymer film contains active groups, so that different functional substances and target molecules can be combined, such as metal atoms, metal oxide nano particles, graphene oxide, functional living cells, proteins, enzymes, polypeptides, metal binding polypeptides, fluorescent dyes, antibodies, anti-antibodies, haptens, immunogens, DNA, oligonucleotides, growth factors, drug macromolecules, ligands, amino and/or sulfhydryl and/or aldehyde and/or carboxyl terminated polyethylene glycol, carbohydrate substances, fatty amines and other substances containing amino and/or sulfhydryl and/or aldehyde and/or carboxyl in the structure, and the like.
The copolymer film and the multilayer film prepared by the invention have rich types of selectable film forming monomers, and can be designed and prepared into films with rich and various components and molecular structures, so that the composition, structure, performance and service performance of the copolymer film are determined, and the copolymer film can be selected according to the application requirements.
The copolymer film prepared by selecting proper film-forming monomers is combined with quaternary ammonium salt molecules to realize antibacterial effect.
The copolymer film can construct supermolecular functional materials, assemble two-dimensional and smaller structural materials and construct multilayer functional polymer materials; the bound or supported catalyst becomes a reactive polymer.
According to embodiments of the present invention, the copolymer membrane may be used for catalysis of biological and chemical reactions by incorporating catalysts, including biological enzymes, chemical catalysts.
The copolymer film prepared by selecting a suitable film-forming monomer can be connected with dye molecules to express corresponding colors.
According to an embodiment of the present invention, the copolymer film may deposit inorganic substances (silver, hydroxyapatite, calcium carbonate, silica, etc.) on the surface thereof to form a nanolayer, and thus an organic-inorganic derivative film may be prepared. For example, a metal modified vesicle is prepared by adding the vesicle to a metal salt solution.
According to an embodiment of the invention, the metal salt includes, but is not limited to, silver nitrate, chloroauric acid, chloroplatinic acid, or palladium nitrate.
Metal cations are reduced from the metal salt solution and deposited on the surface of the copolymer film, thereby allowing electroless metallization of the film surface.
In addition, the copolymer film is prepared, and meanwhile, copolymer particles are generated in the water phase, and the copolymer particles contain high-density amino, quinone, phenolic hydroxyl and other active groups, so that the copolymer particles have the characteristic of re-reaction, and can be added into plastics as fillers to improve the performance of the plastics.
Advantageous effects
The copolymer film and the enzymatic self-assembly synthesis method thereof provided by the invention have the advantages that the operation steps are simple, the isolation of air is not needed, the external potential is not needed, the reaction condition is mild, the preparation can be carried out at normal temperature, normal pressure, neutral or near neutral pH, and the required energy consumption is small; the reaction is carried out in liquid, the chemical uniformity is good, the multi-component uniformity can reach the molecular level, and the composition and the structure are uniform; the reaction is easy to control, the repeatability is good, and the structure can be regulated and controlled at the molecular level. The thickness of the copolymer film, the thickness of the deposit/bond on the film, can be controlled individually on the nanometer scale.
The method for preparing the copolymer film has wide selection range of film forming monomers, can use various film forming monomers, can polymerize monomers containing catechol structures, can polymerize monomers with resorcinol structures, monomers with hydroquinone structures and even compounds with high oxidation-reduction potentials such as bisphenols, biphenyls and the like. On one hand, different film-forming monomers are polymerized and crosslinked by laccase catalysis to obtain films with different molecular structures, and the films with different molecular structures have different properties, performances and applications, so that the design of the films has high degree of freedom and strong designability, and the copolymer films with corresponding functions can be selected and prepared according to the application requirements. On the other hand, the surface of the copolymer film prepared by selecting a proper film-forming monomer can be provided with a selected active group, the corresponding activity and property are displayed, and target molecules and/or atoms can be introduced into the surface of the film through secondary/repeated chemical or/and enzymatic reaction, so that the purpose of functional modification of the surface of the film is achieved, the surface of the polymer film has extremely strong designability, and the application range of the film is further expanded.
The copolymer film of the invention has higher crosslinking degree, thus having better mechanical property and good tolerance to water, acid, alkali, organic solvent and heat. Thus, the vesicles prepared by the invention have better stability than vesicles prepared with surfactants and proteins.
Based on the characteristics, the preparation method is easy to upgrade from laboratory to industrialized mass production, has short design time, is easier to start and stop production, can quickly and flexibly replace products on the same production line, is suitable for small-batch production process and distributed manufacturing, has strong adaptability, and has high flexibility of products, production plans and scheduling. It is particularly noteworthy that the base membranes of the present invention are prepared enzymatically, many of which are difficult to achieve chemically.
The invention has the outstanding beneficial effects that the characteristics of the copolymer film can be controlled manually under the condition of more degrees of freedom, and the material meeting the requirements is obtained, so that the invention has great potential in technical application.
Definition and interpretation of terms
Unless otherwise indicated, the radical and term definitions recited in the specification and claims of this application, including as examples, exemplary definitions, preferred definitions, definitions recited in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. Such combinations and combined group definitions and structures of compounds should fall within the scope of the description herein.
The term "film-forming monomer" refers to an organic compound molecule or group of molecules that becomes a copolymer film by polymerization.
The film-forming monomer containing phenolic hydroxyl groups refers to a compound which is composed of one or more hydroxyl groups in the same molecule and is directly bonded to an aromatic ring.
The film-forming monomer containing at least two amino groups refers to a compound which consists of substances containing at least two amino groups in the same molecule.
The low-melting-point alloy is a metal alloy material which has a melting point of 1-96 ℃, is not easy to volatilize, has stable property and does not react with water phase below 120 ℃.
The artificial enzyme is an enzyme mimic with specific catalytic function synthesized by utilizing an organic chemistry and biology method according to the catalytic mechanism of the enzyme and simulating the biological catalytic function of natural enzyme, and comprises metalloenzyme.
The metalloenzyme refers to a conjugated enzyme containing one or more metal ions as prosthetic groups.
The numerical ranges recited in the specification and claims are equivalent to at least each specific integer number recited therein unless otherwise stated. For example, the numerical ranges "1 to 40" correspond to the values of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, which are each integer in the numerical ranges "1 to 10", and 11, 12, 13, 14, 15, and..the numbers of 35, 36, 37, 38, 39, 40, which are each integer in the numerical ranges "11 to 40". It is to be understood that "more" in one, two or more as used herein in describing substituents shall mean an integer of ≡3, such as 3, 4, 5, 6, 7, 8, 9 or 10. Furthermore, when certain numerical ranges are defined as "numbers," it is to be understood that both endpoints of the range, each integer within the range, and each fraction within the range are delineated. For example, a "number of 0 to 10" should be understood to describe not only each integer of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, but also at least the sum of each integer with 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, respectively.
The term "halogen" means fluorine, chlorine, bromine and iodine.
The term "C 1-40 Alkyl "is understood to mean a straight-chain or branched saturated monovalent hydrocarbon radical having from 1 to 40 carbon atoms. For example, "C 1-10 Alkyl "means straight-chain and branched alkyl having 1,2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms," C 1-6 Alkyl "means straight and branched alkyl groups having 1,2, 3, 4, 5 or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-ethylpropyl, 1, 2-dimethylpropyl, neopentyl, 1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 2, 3-dimethylbutyl, 1, 3-dimethylbutyl, or 1, 2-dimethylbutyl, or the like, or an isomer thereof.
The term "C 2-40 Alkenyl "is understood to mean preferably a straight-chain or branched monovalent hydrocarbon radical which contains one or more double bonds and has from 2 to 40 carbon atoms, preferably" C 2-10 Alkenyl groups). "C 2-10 Alkenyl "is understood to mean preferably a straight-chain or branched monovalent hydrocarbon radical which contains one or more double bonds and has 2,3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, for example 2,3, 4, 5 or 6 carbon atoms (i.e.C 2-6 Alkenyl) having 2 or 3 carbon atoms (i.e., C 2-3 Alkenyl). It will be appreciated that where the alkenyl group comprises more than one double bond, the double bonds may be separated from each other or conjugated. The alkenyl is, for example, vinyl, allyl, (E) -2-methylvinyl, (Z) -2-methylvinyl, (E) -but-2-enyl, (Z) -but-2-enyl, (E) -but-1-enyl, (Z) -but-1-enyl, pent-4-enyl, (E) -pent-3-enyl, (Z) -pent-3-enyl, (E) -pent-2-enyl, (E) -pent-1-enyl, (Z) -pent-1-enyl, hex-5-enyl, (E) -hex-4-enyl, (Z) -hex-4-enyl, (E) -hex-3-enyl, (Z) -hex-3-enyl, (E) -hex-2-enyl, (Z) -hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E) -1-methylprop-1-enyl, (Z) -1-methylbut-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut-2-enyl, (E) -2-methylbut-2-enyl, (Z) -2-methylbut-2-enyl, (E) -1-methylbut-2-enyl, (Z) -1-methylbut-2-enyl, (E) -3-methylbut-1-enyl, (Z) -3-methylbut-1-enyl, (E) -2-methylbut-1-enyl, (Z) -2-methylbut-1-enyl, (E) -1-methylbut-1-enyl, (Z) -1-methylbut-1-enyl, 1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.
The term "C 2-40 Alkynyl "is understood to mean a monovalent hydrocarbon radical, directly or branched, containing one or more triple bonds and having from 2 to 40 carbon atoms, preferably" C 2-10 Alkynyl groups. The term "C 2-10 Alkynyl "is understood to mean preferably a straight-chain or branched monovalent hydrocarbon radical which contains one or more triple bonds and has 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms,for example, having 2, 3, 4, 5 or 6 carbon atoms (i.e., "C 2-6 Alkynyl ") having 2 or 3 carbon atoms (" C 2-3 Alkynyl "). The alkynyl group is, for example, ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hex-4-ynyl, hex-5-ynyl, 1-methylpropan-2-ynyl, 2-methylbutan-3-ynyl, 1-methylbutan-2-ynyl, 3-methylbutan-1-ynyl, 1-ethylpropan-2-ynyl 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpent-4-ynyl, 2-methylpent-3-ynyl, 1-methylpent-3-ynyl, 4-methylpent-2-ynyl, 1-methylpent-2-ynyl, 4-methylpent-1-ynyl, 3-methylpent-1-ynyl, 2-ethylbut-3-ynyl, 1-ethylbut-2-ynyl, 1-propylprop-2-ynyl, 1-isopropylprop-2-ynyl, 2-dimethylbbut-3-ynyl, 1, 1-dimethylbut-3-ynyl, 1-dimethylbut-2-ynyl or 3, 3-dimethylbut-1-ynyl. In particular, the alkynyl group is ethynyl, prop-1-ynyl or prop-2-ynyl.
The term "C 3-40 Cycloalkyl "is understood to mean a saturated monovalent monocyclic, bicyclic hydrocarbon ring or bridged cycloalkane having 3 to 40 carbon atoms, preferably" C 3-10 Cycloalkyl groups). The term "C 3-10 Cycloalkyl "is understood to mean a saturated monovalent monocyclic, bicyclic hydrocarbon ring or bridged cycloalkane having 3,4, 5,6, 7, 8, 9 or 10 carbon atoms. The C is 3-10 Cycloalkyl may be a monocyclic hydrocarbon group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon group such as a decalin ring.
The term "3-to 20-membered heterocyclic group" means a saturated monovalent monocyclic, bicyclic hydrocarbon ring or bridged cycloalkane containing 1 to 5 non-aromatic cyclic groups having 3 to 20 (e.g., 3,4, 5,6, 7, 8, 9, 10, etc. atoms) total ring atoms independently selected from N, O and S heteroatoms, preferably "3-to 10-membered heterocyclic groups". The term "3-to 10-membered heterocyclyl" means a saturated monovalent monocyclic, bicyclic hydrocarbon ring or bridged cycloalkane comprising 1 to 5, preferably 1 to 3 heteroatoms selected from N, O and S. The heterocyclic group may be attached to the remainder of the molecule through any of the carbon atoms or a nitrogen atom, if present. In particular, the heterocyclic groups may include, but are not limited to: 4-membered rings such as azetidinyl, oxetanyl; a 5-membered ring such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6 membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl; or a 7-membered ring such as diazepanyl. Optionally, the heterocyclyl may be benzo-fused. The heterocyclyl may be bicyclic, such as, but not limited to, a 5,5 membered ring, such as hexahydrocyclopenta [ c ] pyrrol-2 (1H) -yl ring, or a 5,6 membered bicyclic ring, such as hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl ring. The nitrogen atom-containing ring may be partially unsaturated, i.e., it may contain one or more double bonds, such as, but not limited to, 2, 5-dihydro-1H-pyrrolyl, 4H- [1,3,4] thiadiazinyl, 4, 5-dihydro-oxazolyl, or 4H- [1,4] thiazide, or it may be benzo-fused, such as, but not limited to, dihydroisoquinolinyl. According to the invention, the heterocyclic group is non-aromatic. When the 3-20 membered heterocyclic group is linked to other groups to form the compound of the present invention, the carbon atom on the 3-20 membered heterocyclic group may be linked to other groups, or the heterocyclic atom on the 3-20 membered heterocyclic ring may be linked to other groups. For example, when the 3-to 20-membered heterocyclic group is selected from piperazinyl, it may be that the nitrogen atom on the piperazinyl group is attached to other groups. Or when the 3-to 20-membered heterocyclic group is selected from piperidyl, it may be that the nitrogen atom on the piperidyl ring and the carbon atom at the para position thereof are linked to other groups.
The term "C 6-20 Aryl "is understood to mean preferably a mono-, bi-or tricyclic hydrocarbon ring, preferably" C ", of monovalent aromatic or partly aromatic nature having from 6 to 20 carbon atoms 6-14 Aryl group). The term "C 6-14 Aryl "is understood to mean preferably a mono-, bi-or tricyclic hydrocarbon ring (" C ") having a monovalent aromatic or partially aromatic character of 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms 6-14 Aryl), in particular a ring having 6 carbon atoms ("C) 6 Aryl')Such as phenyl; or biphenyl, or a ring having 9 carbon atoms ("C 9 Aryl "), e.g. indanyl or indenyl, or a ring having 10 carbon atoms (" C 10 Aryl "), such as tetralin, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms (" C " 13 Aryl "), e.g. fluorenyl, or a ring having 14 carbon atoms (" C) 14 Aryl "), such as anthracenyl. When said C 6-20 When aryl is substituted, it may be mono-substituted or poly-substituted. The substitution site is not limited, and may be, for example, ortho, para or meta substitution.
The term "5-to 20-membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic or tricyclic aromatic ring systems: having 5 to 20 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S, for example "5 to 14 membered heteroaryl". The term "5-to 14-membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic or tricyclic aromatic ring systems: having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and which contain 1 to 5, preferably 1 to 3 heteroatoms each independently selected from N, O and S and which may additionally be benzo-fused in each case. In particular, the heteroaryl group is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, thia-4H-pyrazolyl and the like and their benzo derivatives, such as benzofuryl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazole, indazolyl, indolyl, isoindolyl and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and the like, and their benzo derivatives, such as quinolinyl, quinazolinyl, isoquinolinyl, and the like; or an axcinyl group, an indolizinyl group, a purinyl group, etc., and their benzo derivatives; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and the like. When the 5-to 20-membered heteroaryl is linked to other groups to form the compounds of the invention, the carbon atom on the 5-to 20-membered heteroaryl ring may be linked to other groups, or the heteroatom on the 5-to 20-membered heteroaryl ring may be linked to other groups. When the 5-to 20-membered heteroaryl group is substituted, it may be mono-substituted or poly-substituted. And, the substitution site thereof is not limited, and for example, hydrogen attached to a carbon atom on a heteroaryl ring may be substituted, or hydrogen attached to a heteroatom on a heteroaryl ring may be substituted.
As used herein, the term "derivative" refers to a chemical substance that is structurally related to another, which is a "native" substance, and may also be referred to as a "parent" compound. The "derivatives" may be manufactured in one or more steps from a structurally related parent compound.
Unless otherwise indicated, heterocyclyl, heteroaryl or heteroarylene include all possible isomeric forms thereof, e.g. positional isomers thereof. Thus, for some illustrative non-limiting examples, forms that may include substitution at one, two, or more of its 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-positions, etc. (if present) or bonding to other groups include pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, and pyridin-4-yl; thienyl or thienylene include thiophen-2-yl, thienylene-2-yl, thiophen-3-yl and thienylene-3-yl; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.
The term "oxo" refers to the substitution of a carbon atom, nitrogen atom or sulfur atom in a substituent with an oxo group (=o) formed after oxidation.
By polyphenol is meant a natural, synthetic and semisynthetic organic compound characterized by the presence of a large number of repeating phenolic building blocks. Polyphenols are generally compounds having a molecular weight of 400-4000 Da, greater than or equal to 5 phenolic hydroxyl groups.
The term "enhancer of enzymes" refers to diffusible molecules activated by oxidase and diffusing from active sites on the enzyme to sensitive structures, which are small organic compounds of low redox potential that act as carriers for electron transfer during laccase catalytic oxidation.
The multilayer film is a film composed of at least one layer of copolymer film. In order to distinguish the film that is initially formed from a film that is prepared by continuing the reaction two or more times on its basis, the copolymer film that is initially formed is referred to as a "base film" in the present invention. The base film is a copolymer film prepared by two or more chemical or/and enzymatic reactions, referred to as a "derivative film". The new film layer can be generated on one side or two sides of the basic film through enzymatic polymerization, the chemical composition and structure of the new film layer can be different from those of the basic film, and the new film layer can be continuously generated. The base film and the new film layer together form a "multilayer film". The derivative film continues to form a new polymer film, known as a "derivative multilayer film".
Detailed Description
The technical scheme of the invention will be further described in detail below with reference to specific embodiments. It is to be understood that the following examples are illustrative only and are not to be construed as limiting the scope of the invention. All techniques implemented based on the above description of the invention are intended to be included within the scope of the invention.
The Polyethyleneimine (PEI) in the present invention has an average relative molecular weight (average relative molecular) of 600, manufactured by Shanghai microphone Biochemical technology Co., ltd. The paraffin is refined paraffin with a melting point of 58-60 ℃ and is manufactured by Jiangsu Shitai laboratory instruments Co. Unless otherwise indicated, the starting materials and reagents used in the following examples were either commercially available or may be prepared by known methods.
Definition of laccase enzyme activity: the amount of enzyme required to oxidize 1. Mu. Mol of 2,2' -Azino-bis- (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) per minute was 1 enzyme activity unit (U).
Definition of manganese peroxidase enzyme activity: the amount of enzyme required to oxidize 1. Mu. Mol of ABTS per minute was 1 enzyme activity unit (U).
Definition of horseradish peroxidase enzymatic activity: the amount of enzyme required to oxidize 1. Mu. Mol of ABTS per minute was 1 enzyme activity unit (U).
Definition of lignin peroxidase enzyme activity: the amount of enzyme required to oxidize 1. Mu. Mol of ABTS per minute was 1 enzyme activity unit (U).
Definition of soybean peroxidase enzymatic activity: the amount of enzyme required to oxidize 1. Mu. Mol of ABTS per minute was 1 enzyme activity unit (U).
Definition of chloroperoxidase enzyme activity: the amount of enzyme required for oxidizing 1. Mu. Mol of 2-chloro-5, 5-dimethyl-1, 3-cyclohexanedione per minute to produce 2, 2-dichloro-5, 5-dimethyl-1, 3-cyclohexanedione was 1 enzyme activity unit (U).
Definition of monophenol monooxidase enzyme activity: the amount of enzyme required to oxidize 1. Mu. Mol of L-DOPA (L-DOPA) per minute was 1 enzyme activity unit (U).
Definition of catechol oxidase enzyme activity: the amount of enzyme required to oxidize 1. Mu. Mol of catechol per minute was 1 enzyme activity unit (U).
Definition of bilirubin oxidase enzyme activity: the amount of enzyme required to oxidize 1. Mu. Mol of ABTS per minute was 1 enzyme activity unit (U).
Example 1: preparation of laccase
(1) Liquid seed medium: 30g/L corn flour, 15g/L bean cake flour, 54U/L alpha-amylase and NaH 2 PO 4 2.6g/L，KCl 2.25g/L，MgSO 4 ·7H 2 O1.5 g/L, water in balance, pH 6.0,0.1MPa for 25min.
Fermentation medium: 48g/L fructose, 12g/L corn meal, 9g/L soybean peptone, (NH) 4 ) 2 SO 4 0.5g/L，KCl 1g/L，NaH 2 PO 4 1.8g/L，MgSO 4 ·7H 2 O 0.25g/L，CuSO 4 ·5H 2 O 1mmol/L，VB 1 0.03g/L Tween-80.5 g/L, vanillin 1mmol/L, water balance, pH 6.5,0.1MPa sterilization 25min.
(2) Culturing liquid seeds: taking inclined plane lawn of winter-growing porous fungus (Polyporus brumalis) (strain with preservation number CCTCC NO: M2020809) about 3cm 2 Inoculating to 500mL triangular flask, and culturing in 150mL liquid seed culture medium at 30deg.C and 150rpm for 4 days to obtain seed solution.
Shake flask fermentation culture: inoculating the seed solution to a fermentation culture medium according to the inoculum size of 6% of the volume ratio, wherein the liquid loading amount of the fermentation culture medium in a 500mL triangular flask is 150mL, carrying out shake culture at 30 ℃, and the rotation speed of a shaking table after fermentation for 1-3 days is 150rpm and the rotation speed of the shaking table after fermentation for 3 days is 200rpm.
(3) Preparation of crude enzyme solution: the fermentation broth is centrifuged for 10min at 10000 Xg at 4 ℃, and the supernatant is the crude enzyme solution of laccase.
(4) Purification of laccase:
taking 100mL of coarse enzyme solution of laccase, slowly adding ammonium sulfate in stages while stirring to ensure that the concentration of the ammonium sulfate solution gradually reaches 60%, centrifuging at 12000 Xg for 10min at 4 ℃ after full precipitation, collecting precipitate, and dissolving the precipitate in a proper volume of pH 7.0 and 0.02mol/L citric acid-disodium hydrogen phosphate buffer solution to obtain salting-out solution. And (3) placing the salting-out solution in the same buffer solution for dialysis overnight, and replacing the dialysis solution every 10 hours until the conductivity and the pH value of the liquid inside and outside the dialysis bag are the same, so as to obtain the dialysis solution of laccase.
The column packed with DEAE-Sepharose Fast Flow ion exchange medium was equilibrated with citric acid-disodium hydrogen phosphate buffer solution (solution A) at pH 7.0,0.02 mol/L. The enzyme solution obtained by dialysis was filtered through a filter membrane having a pore size of 0.22. Mu.m, and then was applied at a flow rate of 2mL/min in an amount of 10mL. After sample loading, the sample is balanced by the solution A to be at the base line zero, then continuous gradient elution is carried out by the buffer solution (solution B) of citric acid and disodium hydrogen phosphate containing NaCl of 0-0.5 mol/L respectively, the flow rate is 1mL/min, the absorbance at the wavelength of 280nm is recorded, active components are collected for standby, and each tube is collected for 2mL. And (3) measuring laccase activity and protein concentration of the collected sample, combining eluents with laccase activity to obtain laccase purified by a chromatographic column (called pure enzyme solution in the invention), and storing at-20 ℃ for later use.
Example 2: preparation of copolymer film by enzymatic polymerization at liquid paraffin-water solution interface
(1) Laccase catalyzed preparation of a ***e-arginine copolymer film:
0.050g of caffeine, 0.050g of arginine and 5.0mg of vanillin are dissolved in 35mL of a mixed solvent of 0.05mol/L sodium succinate-succinic acid buffer solution (pH 2.0) and 15mL of acetone, 7.5U of laccase is added, and the mixture is fully and uniformly mixed to obtain a laccase catalytic system (aqueous phase). The aqueous phase was covered with 3mm thick liquid paraffin, and left to stand in an incubator (ZSD-1090, china) at 30℃for 3 hours, whereby a "***e-arginine copolymer film" was formed at the interface between the liquid paraffin and the aqueous phase. The transfer process of the "corin-arginine copolymer membrane" is as follows: sucking out most liquid paraffin on the upper side of the 'corin-arginine copolymer membrane' and water phase on the lower side of the 'corin-arginine copolymer membrane' by using a syringe, and injecting distilled water on the lower side of the 'corin-arginine copolymer membrane', so that the liquid suction-liquid injection-liquid suction operation is repeated for a plurality of times; the coverslip was inserted into the water phase under the membrane, carefully moved under the "corin-arginine copolymer membrane" and gently fished out, and the residual liquid paraffin on the membrane surface was sucked off with filter paper and dried at room temperature.
(2) tectorigenin-PEI copolymer membrane prepared by laccase catalysis:
A laccase catalytic system (aqueous phase) was obtained by dissolving 0.250g tectorigenin, 0.250g PEI, 12.5mg ABTS in 175mL of a mixed solvent of 0.05mol/L potassium hydrogen phthalate-sodium hydroxide buffer solution (pH 5.0) and 75mL of ethanol, adding laccase 50U, and mixing uniformly. The aqueous phase was covered with 5mm thick liquid paraffin, and left to stand in an oven (Zhiyeng, ZFD-5090, china) at 60℃for 8 hours, whereby an "tectorigenin-PEI copolymer film" was formed at the interface between the liquid paraffin and the aqueous phase.
The transfer method of the film comprises the following steps: the bottom aqueous phase was slowly drained, allowing the film to fall onto the substrate surface at the bottom of the container, removing the upper liquid wax layer.
(3) Preparation of rhein-2, 4-diamino-6-methyl-1, 3, 5-triazine copolymer film catalyzed by laccase:
0.125g rhein, 0.125g 2, 4-diamino-6-methyl-1, 3, 5-triazine and 20mg syringaldehyde are dissolved in 175mL mixed solvent of 0.05mol/L sodium acetate-acetic acid buffer solution (pH 4.0) and 75mL methanol, laccase 5U is added, and the mixture is uniformly mixed to obtain a laccase catalytic system (water phase). Liquid paraffin with the thickness of 5mm is covered on the water phase, and the mixture is placed in an oven (Zhiyeng, ZFD-5090, china) at the temperature of 50 ℃ for reaction for 10 hours. The interface between the liquid paraffin and the aqueous phase forms a 'rhein-2, 4-diamino-6-methyl-1, 3, 5-triazine copolymer film'.
(4) Preparation of 1, 6-dihydroxynaphthalene-diethylenetriamine copolymer film by manganese peroxidase catalysis:
dissolving 0.125g of 1, 6-dihydroxynaphthalene and 0.250g of diethylenetriamine in 200mL of a mixed solvent of 0.05mol/L disodium hydrogen phosphate-citric acid buffer solution (pH 4.0) and 50mL of acetone, adding 12U of manganese peroxidase, 40 mu L of 5% hydrogen peroxide and 0.1mol/L MnSO 4 3.5mL, and mixing to obtain a manganese peroxidase catalytic system (water phase). Liquid paraffin of 5mm thickness was covered on the surface of the aqueous phase with a pipette, and left to stand in an incubator (zhiyeng, ZSD-1090, china) at 37 ℃ for 10 hours, during which 5% hydrogen peroxide was added to the reaction system several times, 40 μl each time. The interface between the liquid paraffin and the aqueous phase forms a "1, 6-dihydroxynaphthalene-diethylenetriamine copolymer film".
(5) Manganese peroxidase catalysis is used for preparing shikonin-lysine copolymer film:
0.125g shikonin and 0.125g lysine are dissolved in 175mL of mixed solvent of 0.05mol/L disodium hydrogen phosphate-citric acid buffer solution (pH 3.0) and 75mL of ethanol, and manganese peroxidase 10U, 5% hydrogen peroxide 40 mu L and 0.1mol/L MnSO are added 4 3.5mL, and mixing to obtain a manganese peroxidase catalytic system (water phase). The surface of the manganese peroxidase catalytic system is covered with liquid paraffin with the thickness of 5mm by using a straw, and the liquid paraffin is placed in a baking oven (Zhiyeng, ZFD-5090, china) at the temperature of 45 ℃ for 6 hours, and 5% hydrogen peroxide is added to the reaction system for multiple times, and 40 mu L of hydrogen peroxide is added each time. The interface between the liquid paraffin and the aqueous phase forms a "shikonin-lysine copolymer film".
(6) Preparation of 3-methyl salicylic acid-3-nitro-1, 2-phenylenediamine copolymer film catalyzed by manganese peroxidase:
0.010g of 3-methyl salicylic acid, 0.010g of 3-nitro-1, 2-phenylenediamine was dissolved in 50mL of 0.05mol/L sodium succinate-succinic acid buffer solution (pH 5.0), and manganese peroxidase 0.05U, 5% hydrogen peroxide 8. Mu.L and 0.1mol/L MnSO were added 4 0.7mL, and the manganese peroxidase catalytic system (water phase) is obtained after uniform mixing. The surface of the manganese peroxidase catalytic system is covered with liquid paraffin with the thickness of 5mm by using a straw, and the liquid paraffin is placed in an incubator (ZSD-1090, china) at the temperature of 20 ℃ for reaction for 12 hours, and 5% hydrogen peroxide is added to the reaction system for multiple times during the reaction, and 8 mu L of the liquid paraffin is added each time. Formation of "3-methyl salicylic acid-3-nitro-1, 2-phenylenediamine copolymer film" at the interface of liquid paraffin and aqueous phase”。
(7) Preparation of a galloblue-2, 4, 6-triaminopyrimidine copolymer membrane by manganese peroxidase catalysis:
0.250g of galloblue and 0.250g of 2,4, 6-triaminopyrimidine were dissolved in 250mL of 0.05mol/L potassium hydrogen phthalate-sodium hydroxide buffer solution (pH 5.0), and manganese peroxidase 10U, 5% hydrogen peroxide 40. Mu.L and 0.1mol/L MnSO were added 4 3.5mL, and mixing to obtain a manganese peroxidase catalytic system (water phase). The surface of the manganese peroxidase catalytic system is covered with liquid paraffin with the thickness of 5mm by using a straw, and the liquid paraffin is placed in a baking oven (Zhiyeng, ZFD-5090, china) at the temperature of 40 ℃ for reaction for 15 hours, and 5 percent of hydrogen peroxide is added into the reaction system for multiple times, and 40 mu L of hydrogen peroxide is added each time. The interface between the liquid paraffin and the aqueous phase forms a "galloblue-2, 4, 6-triaminopyrimidine copolymer film".
(8) Preparing a 4,4' -dihydroxybiphenyl-2, 3-diaminonaphthalene copolymer film by horseradish peroxidase catalysis:
0.125g of 4,4' -dihydroxybiphenyl, 0.125g of 2, 3-diaminonaphthalene are dissolved in 215mL of a mixed solvent of 0.05mol/L glycine-sodium hydroxide buffer solution (pH 10.0) and 35mL of acetone, 6U of horseradish peroxidase and 40 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain a horseradish peroxidase catalytic system (water phase). Liquid paraffin with the thickness of 10mm is covered on the surface of a horseradish peroxidase catalytic system by a straw, and the mixture is kept stand for 24 hours in a baking oven (Zhiyeng, ZFD-5090, china) at the temperature of 40 ℃, and 5% hydrogen peroxide is added to the reaction system for multiple times, and 40 mu L of hydrogen peroxide is added each time. The interface between the liquid paraffin and the aqueous phase forms a "4,4' -dihydroxybiphenyl-2, 3-diaminonaphthalene copolymer film".
(9) Preparing bisphenol A-proflavone copolymer membrane by horseradish peroxidase catalysis:
0.250g bisphenol A, 0.250g pre-flavin are dissolved in 175mL mixed solvent of 0.1mol/L Tris-hydrochloric acid buffer solution (pH 8.0) and 75mL ethanol, and a proper amount of horseradish peroxidase 3U and 5% hydrogen peroxide 40 mu L are added and uniformly mixed to obtain a horseradish peroxidase catalytic system (water phase). Liquid paraffin with the thickness of 5mm is covered on the surface of a horseradish peroxidase catalytic system by a straw, and the mixture is kept stand for 5 hours in a baking oven (Zhiyeng, ZFD-5090, china) at the temperature of 45 ℃, and 5% hydrogen peroxide is added to the reaction system for multiple times, and 40 mu L of hydrogen peroxide is added each time. The interface between the liquid paraffin and the aqueous phase forms a bisphenol A-procxanthin copolymer film.
(10) Preparation of 1,8, 9-trihydroxy anthracene-4, 4' -diaminodiphenyl sulfone copolymer membrane catalyzed by horseradish peroxidase:
0.040g of 1,8, 9-trihydroxyanthracene, 0.060g of 4,4' -diaminodiphenyl sulfone are dissolved in 140mL of a mixed solvent of 0.05mol/L disodium hydrogen phosphate-citric acid buffer solution (pH 6.0) and 60mL of ethanol, 12U of horseradish peroxidase and 40 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain a horseradish peroxidase catalytic system (water phase). Liquid paraffin with the thickness of 5mm is covered on the surface of a horseradish peroxidase catalytic system by a straw, and the mixture is kept stand for 17 hours in a baking oven (Zhiyeng, ZFD-5090, china) at the temperature of 45 ℃, and 5% hydrogen peroxide is added to the reaction system for multiple times, and 40 mu L of hydrogen peroxide is added each time. The interface between the liquid paraffin and the aqueous phase forms a "1,8, 9-trihydroxyanthracene-4, 4' -diaminodiphenyl sulfone copolymer film".
(11) Preparation of 4',6, 7-dihydroxyisoflavone-auxiliary product red copolymer film by lignin peroxidase catalysis:
0.750g of 4',6, 7-trihydroxyisoflavone, 1.500g of parafuchsin are dissolved in 175mL of a mixed solvent of 0.05mol/L sodium citrate-citric acid buffer solution (pH 5.0) and 75mL of ethanol, 1.5U of lignin peroxidase and 40 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain a lignin peroxide catalytic system (water phase). The lignin peroxide catalytic system surface is covered with liquid paraffin with the thickness of 5mm by a straw, and the liquid paraffin is kept stand in a 45 ℃ oven (Zhiyeng, ZFD-5090, china) for reaction for 12 hours, and 5% hydrogen peroxide is added to the reaction system for multiple times, and 40 mu L of hydrogen peroxide is added each time. The interface between the liquid paraffin and the aqueous phase forms a "4',6, 7-trihydroxyisoflavone-subsidiary red copolymer film".
(12) Catalytic preparation of a geraniin-1, 6-hexamethylenediamine copolymer membrane with lignin peroxidase:
0.025g of geraniin, 0.050g of 1, 6-hexamethylenediamine are dissolved in 225mL of mixed solvent of trisodium phosphate-phosphate buffer solution (pH 4.0) and 25mL of dimethyl sulfoxide, 6U of lignin peroxidase and 40 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain a lignin peroxide catalytic system (water phase). The surface of lignin peroxide catalytic system is covered with liquid paraffin with the thickness of 5mm by using a straw, and the liquid paraffin is placed in a incubator (Zhiyeng, ZSD-1090, china) at the temperature of 30 ℃ for reaction for 8 hours, and 5% hydrogen peroxide is added into the reaction system for multiple times, and 40 mu L of hydrogen peroxide is added each time. The interface between the liquid paraffin and the aqueous phase forms a 'geraniin-1, 6-hexamethylenediamine copolymer film'.
(13) Preparation of resveratrol-5, 6-diamino-1, 3-dimethyluracil copolymer membrane by soybean peroxidase catalysis:
0.025g of resveratrol, 0.025g of 5, 6-diamino-1, 3-dimethyluracil is dissolved in 35mL of a mixed solvent of 0.05mol/L trisodium phosphate-phosphoric acid buffer solution (pH 6.0) and 15mL of ethanol, and 0.6U of soybean peroxidase and 8 mu L of 5% hydrogen peroxide are added and uniformly mixed to obtain a soybean peroxidase catalytic system (water phase). The surface of the soybean peroxidase catalytic system is covered with liquid paraffin with the thickness of 5mm by using a suction pipe, and the liquid paraffin is placed in a baking oven (Zhiyeng, ZFD-5090, china) at 65 ℃ for reaction for 5 hours, and 5% hydrogen peroxide is added to the reaction system for multiple times, and 8 mu L of the liquid paraffin is added each time. The interface between the liquid paraffin and the water phase forms a resveratrol-5, 6-diamino-1, 3-dimethyluracil copolymer film.
(14) Preparation of esculetin-melamine-1, 4-diaminoanthraquinone copolymer film catalyzed by soybean peroxidase:
0.075g esculetin, 0.125g melamine, 0.075g 1, 4-diaminoanthraquinone are dissolved in 175mL mixed solvent of 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 5.0) and 75mL ethanol, 15U of soybean peroxidase and 40 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain a soybean peroxidase catalytic system (water phase). The surface of the soybean peroxidase catalytic system is covered with liquid paraffin with the thickness of 5mm by using a suction pipe, and the mixture is placed in a baking oven (Zhiyeng, ZFD-5090, china) at the temperature of 75 ℃ for reaction for 4 hours, and 5% hydrogen peroxide is added to the reaction system for multiple times, and 40 mu L of hydrogen peroxide is added each time. The interface between the liquid paraffin and the aqueous phase forms a "esculetin-melamine-1, 4-diaminoanthraquinone copolymer film".
(15) Preparation of ellagic acid-PEI copolymer film catalyzed by chloroperoxidase:
0.025g ellagic acid, 0.050g PEI were dissolved in 250mL of 0.05mol/L sodium citrate-citric acid buffer (pH 3.0), 11.25U of chloroperoxidase and 40. Mu.L of 5% hydrogen peroxide were added and mixed well to give a chloroperoxidase catalytic system (aqueous phase). Liquid paraffin with the thickness of 5mm is covered on the surface of a chloroperoxidase catalytic system by a suction pipe, and the liquid paraffin is placed in an incubator (ZSD-1090, china) at 35 ℃ for reaction for 4 hours, and 5% hydrogen peroxide is added to the reaction system for multiple times during the reaction, and 40 mu L of the liquid paraffin is added each time. An "ellagic acid-PEI copolymer film" is formed at the interface of the liquid paraffin and the aqueous phase.
(16) Preparing a Jinsong biflavone-spermine copolymer membrane by using chloroperoxidase as a catalyst:
0.050g of pinus koraiensis biflavone and 0.100g of spermine are dissolved in 70mL of a mixed solvent of 0.05mol/L trisodium phosphate-phosphoric acid buffer solution (pH 3.0) and 30mL of ethanol, 3U of chloroperoxidase and 16 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain a chloroperoxidase catalytic system (water phase). Liquid paraffin with the thickness of 5mm is covered on the surface of a chloroperoxidase catalytic system by a suction pipe, and the liquid paraffin is kept stand in a baking oven (Zhiyeng, ZFD-5090, china) at the temperature of 40 ℃ for reaction for 6 hours, and 5% hydrogen peroxide is added to the reaction system for multiple times, and 16 mu L of hydrogen peroxide is added each time. The interface between the liquid paraffin and the aqueous phase forms a 'Jinsong biflavone-spermine copolymer film'.
(17) Preparing a colchicoside-2, 4-diaminoanisole copolymer membrane by using chloroperoxidase as a catalyst:
0.075g of Narcin, 0.125g of 2, 4-diaminoanisole are dissolved in 200mL of a mixed solvent of 0.05mol/L sodium malonate-malonic acid buffer solution (pH 4.0) and 50mL of ethanol, 0.75U of chloroperoxidase and 40 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain a chloroperoxidase catalytic system (water phase). Liquid paraffin with the thickness of 5mm is covered on the surface of a chloroperoxidase catalytic system by a suction pipe, and the liquid paraffin is placed in an incubator (ZSD-1090, china) at the temperature of 10 ℃ for reaction for 5 hours, and 5% hydrogen peroxide is added into the reaction system for multiple times during the reaction, and 40 mu L of the liquid paraffin is added each time. The interface between the liquid paraffin and the aqueous phase forms a "colchicoside-2, 4-diaminoanisole copolymer film".
(18) Preparing a tyrosine-o-tolidine copolymer membrane by catalysis of monophenol monooxidase:
0.075g of tyrosine and 0.050g of o-tolidine are dissolved in 250mL of 0.05mol/L disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution (pH 6.5), 150U of monophenol monooxidase is added, and the mixture is uniformly mixed to obtain a monophenol monooxidase catalytic system (water phase). Liquid paraffin with the thickness of 5mm is covered on the surface of a monophenol monooxidase catalytic system by a straw, and the liquid paraffin is placed in an incubator (Zhiyeng, ZSD-1090, china) at the temperature of 20 ℃ for reaction for 36h. The interface between the liquid paraffin and the aqueous phase forms a 'tyrosine-o-tolidine copolymer film'.
(19) Catechol-urea copolymer membranes are prepared catalytically with catechol oxidase:
7.500g catechol and 5.000g urea were dissolved in 250mL of a 0.05mol/L sodium succinate-succinic acid buffer solution (pH 4.0), and catechol oxidase 0.75U was added and mixed well to obtain a catechol oxidase catalyst system (aqueous phase). Covering liquid paraffin with 5mm thickness on the surface of catechol oxidase catalytic system with a straw, standing in oven (ZFD-5090, china) at 50deg.C for reaction for 8 hr. The "catechol-urea copolymer film" is formed at the interface of the liquid paraffin and the aqueous phase.
(20) Preparation of hesperetin-6-hydroxy-2, 4, 5-triaminopyrimidine copolymer membrane catalyzed by bilirubin oxidase:
0.020g of hesperetin, 0.020g of 6-hydroxy-2, 4, 5-triaminopyrimidine are dissolved in 140mL of a mixed solvent of 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 5.0) and 60mL of ethanol, and 100U of bilirubin oxidase is added and uniformly mixed to obtain a bilirubin oxidase catalytic system (water phase). Liquid paraffin with the thickness of 5mm is covered on the surface of a bilirubin oxidase catalytic system by a straw, and the liquid paraffin is placed in a 55 ℃ oven (Zhiyeng, ZFD-5090, china) for reaction for 8 hours. The interface between the liquid paraffin and the aqueous phase forms a 'hesperetin-6-hydroxy-2, 4, 5-triaminopyrimidine copolymer film'.
Example 3: preparation of copolymer film by enzymatic polymerization at aqueous solution-carbon tetrachloride interface
(1) Preparation of 1, 3-dihydroxynaphthalene-kanamycin copolymer film catalyzed by laccase:
0.25g of 1, 3-dihydroxynaphthalene and 0.25g of kanamycin are dissolved in 50mL of 0.05mol/L sodium acetate-acetic acid buffer solution (pH 4.0), 0.6U of laccase is added, and the mixture is uniformly mixed to obtain a laccase catalytic system (aqueous phase). Firstly, adding CCl with the thickness of 55mm into a container 4 (liquid phase which is mutually insoluble in water) and then to the containerSlowly adding water phase, and floating in CCl 4 On the two phases, a liquid-liquid interface is formed between the two phases. Standing in 35℃incubator (ZSD-1090, china) for reaction for 12h. A "1, 3-dihydroxynaphthalene-kanamycin copolymer film" (gas-liquid interfacial film) was formed on the upper surface of the aqueous phase (gas-liquid interface of aqueous phase and air), while at the same time, in the aqueous phase and CCl 4 The liquid-liquid interface of (a) forms a "1, 3-dihydroxynaphthalene-kanamycin copolymer film" (liquid-liquid interface film). The transfer of two interfacial films to coverslips: (1) the cover glass is first inserted into the water phase, carefully moved to the lower part of the 1, 3-dihydroxynaphthalene-kanamycin of the gas-liquid interface, and gently fished out to complete the transfer of the gas-liquid interface film. (2) Sucking out the solution on the upper side of the "1, 3-dihydroxynaphthalene-kanamycin copolymer film" of the liquid-liquid interface by a syringe, re-injecting distilled water on the upper side of the film, repeating the pipetting-liquid-pipetting operation multiple times, and inserting another coverslip into the CCl 4 In the process, the liquid-liquid interface film is gently fished up after being carefully moved below the liquid-liquid interface film, the liquid-liquid interface film is spread on a cover glass in a flat manner, and residual CCl on the lower side of the film is sucked by filter paper 4 And (5) airing at room temperature.
(2) Laccase catalyzed preparation of ferulic acid-2, 3-dimethyl-p-phenylenediamine copolymer film:
0.125g of ferulic acid and 0.125g of 2, 3-dimethyl-p-phenylenediamine are dissolved in 250mL of 0.05mol/L phthalic acid-hydrochloric acid buffer solution (pH 3.0), 0.5U of laccase is added, and the mixture is uniformly mixed to obtain a laccase catalytic system (water phase). Firstly, adding CCl with the thickness of 5mm into a container 4 (liquid phase which is mutually insoluble with water), then slowly adding an aqueous phase to the vessel, the aqueous phase floating in CCl 4 On, aqueous phase and CCl 4 Forming a liquid-liquid interface therebetween. The reaction was allowed to stand at room temperature for 2 hours. Forming a 'ferulic acid-2, 3-dimethyl-p-phenylenediamine copolymer film' on the upper surface of the aqueous phase (gas-liquid interface of the aqueous phase and air), and simultaneously in water, CCl 4 The liquid-liquid interface of the two phases forms a "ferulic acid-2, 3-dimethyl-p-phenylenediamine copolymer film".
(3) Preparation of p-coumaric acid-p-phenylenediamine copolymer membranes catalyzed by manganese peroxidase:
0.125g p-coumaric acid, 0.25g p-phenylenediamine were dissolved in 250mL of 0.05 mol-L tartaric acid-sodium tartrate buffer solution (pH 5.0), manganese peroxidase 0.125U, 5% hydrogen peroxide 40 μL and 0.1mol/LMnSO 4 3.5mL, and mixing to obtain a manganese peroxidase catalytic system (water phase). Firstly, adding CCl with the thickness of 35mm into a container 4 (liquid phase which is mutually insoluble with water) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On, aqueous phase and CCl 4 Forming a liquid-liquid interface therebetween. The reaction mixture was allowed to stand in an incubator (ZSD-1090, china) at 10℃for 36 hours, during which time 5% hydrogen peroxide was added to the aqueous system 40. Mu.L each time. The upper surface of the aqueous phase (the gas-liquid interface of the aqueous phase and air) forms a 'p-coumaric acid-p-phenylenediamine film' on water, CCl simultaneously 4 Form a "p-coumaric acid-p-phenylenediamine copolymer film".
(4) Preparing a 3-methoxy salicylic acid-2, 3-diethyl-p-phenylenediamine copolymer film by horseradish peroxidase catalysis:
0.125g of 3-methoxysalicylic acid and 0.125g of 2, 3-diethyl-p-phenylenediamine are dissolved in 250mL of 0.05mol/L dipotassium hydrogen phosphate-sodium hydroxide buffer solution (pH 6.5), 1.5U of horseradish peroxidase and 40 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain a horseradish peroxidase catalytic system (water phase). Firstly, adding CCl with the thickness of 25mm into a container 4 (liquid phase which is mutually insoluble with water) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On, aqueous phase and CCl 4 Forming a liquid-liquid interface therebetween. The reaction was allowed to stand in an oven (ZFD-5090, china) at 55deg.C for 8 hours, during which time 5% hydrogen peroxide was added to the aqueous phase 40. Mu.L each time. The upper surface of the aqueous phase (gas-liquid interface of the aqueous phase and air) forms "3-methoxy salicylic acid-2, 3-diethyl-p-phenylenediamine" while in water, CCl 4 Form the 3-methoxy salicylic acid-2, 3-diethyl-p-phenylenediamine.
(5) Preparing a 3, 5-diiodosalicylic acid-2-hydroxymethyl-p-phenylenediamine copolymer membrane by lignin peroxidase catalysis:
0.050g of 3, 5-diiodosalicylic acid, 0.100g of 2-hydroxymethyl-p-phenylenediamine were dissolved in 250mL of 0.05mol/L tartaric acid-sodium tartrate buffer solution (pH 3.0), and lignin peroxidase 0.2 was added5U and 40. Mu.L of 5% hydrogen peroxide were mixed to obtain a lignin peroxide catalyst system (aqueous phase). Firstly, adding CCl with the thickness of 25mm into a container 4 (liquid phase which is mutually insoluble with water) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On the water phase and CCl 4 Forming a liquid-liquid interface therebetween. The reaction mixture was allowed to stand in an incubator (ZSD-1090, china) at 37℃for 3 hours, during which time 5% hydrogen peroxide was added to the aqueous phase 40. Mu.L each time. The upper surface of the aqueous phase (gas-liquid interface of the aqueous phase and air) forms a '3, 5-diiodosalicylic acid-2-hydroxymethyl-p-phenylenediamine copolymer film', and simultaneously, the copolymer film is formed on water and CCl 4 The liquid-liquid interface of (a) forms a '3, 5-diiodosalicylic acid-2-hydroxymethyl-p-phenylenediamine copolymer film'.
(6) Preparing gentisic acid-2-hydroxyethoxy-p-phenylenediamine copolymer membrane by lignin peroxidase catalysis:
0.250g gentisic acid and 0.250g 2-hydroxy ethoxy-p-phenylenediamine are dissolved in 250mL of 0.2mol/L disodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution (pH 6.5), 0.5U of lignin peroxidase and 40 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain a lignin peroxide catalytic system (water phase). Firstly, adding CCl with the thickness of 25mm into a container 4 (liquid phase which is mutually insoluble with water) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On the water phase and CCl 4 Form a liquid-liquid interface between the liquid phases of (a). The reaction was allowed to stand in an oven (ZFD-5090, china) at 40℃for 36h, during which time 5% hydrogen peroxide was added to the aqueous phase 40. Mu.L each time. The upper surface of the aqueous phase (the gas-liquid interface of the aqueous phase and air) forms a "gentisic acid-2-hydroxyethoxy-p-phenylenediamine copolymer film" with water, CCl 4 Form a "gentisic acid-2-hydroxyethoxy-p-phenylenediamine copolymer film".
(7) Preparation of para-aminosalicylic acid-m-phenylenediamine copolymer film by soybean peroxidase catalysis:
dissolving 0.250g of p-aminosalicylic acid and 0.250g of m-phenylenediamine in 250mL of 0.05mol/L disodium hydrogen phosphate-citric acid buffer solution (pH 7.0), adding 0.25U of soybean peroxidase and 40 mu L of 5% hydrogen peroxide, and uniformly mixing to obtain the soybean peroxidase catalystSystem (aqueous phase). Firstly, adding CCl with the thickness of 25mm into a container 4 (liquid phase which is mutually insoluble with water) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On the water phase and CCl 4 Forming a liquid-liquid interface therebetween. The reaction was allowed to stand in an incubator (ZSD-1090, china) at 10℃for 24 hours, during which time 5% hydrogen peroxide was added to the aqueous phase 40. Mu.L each time. The upper surface of the aqueous phase (gas-liquid interface of the aqueous phase and air) forms a 'p-aminosalicylic acid-m-phenylenediamine copolymer film', and simultaneously, the p-aminosalicylic acid-m-phenylenediamine copolymer film is formed on water and CCl 4 Form a "para-aminosalicylic acid-meta-phenylenediamine copolymer film".
(8) Preparation of catechol-caffeic acid-3, 4-diaminotoluene copolymer membranes with soybean peroxidase catalysis:
0.025g catechol, 0.025g caffeic acid and 0.050g 3, 4-diaminotoluene were dissolved in 50mL of 0.05mol/L tartaric acid-sodium tartrate buffer solution (pH 4.0), and soybean peroxidase 0.15U and 5% hydrogen peroxide 8. Mu.L were added and mixed to obtain a soybean peroxidase catalyst system (aqueous phase). Firstly, adding CCl with the thickness of 25mm into a container 4 (liquid phase which is mutually insoluble with water) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On the water phase and CCl 4 Forming a liquid-liquid interface therebetween. The reaction mixture was allowed to stand in an incubator (ZSD-1090, china) at 20℃for 6 hours, during which time 5% hydrogen peroxide was added to the aqueous phase 8. Mu.L each time. The upper surface of the aqueous phase (gas-liquid interface of the aqueous phase and air) forms a "catechol-caffeic acid-3, 4-diaminotoluene copolymer film" while in water, CCl 4 Form a "catechol-caffeic acid-3, 4-diaminotoluene copolymer film".
(9) Preparation of syringic acid-3, 4-diaminobenzoic acid copolymer membrane catalyzed by chloroperoxidase:
0.375g of syringic acid and 0.200g of 3, 4-diaminobenzoic acid are dissolved in 250mL of 0.05mol/L sodium acetate-acetic acid buffer solution (pH 3.5), 1.25U of chloroperoxidase and 40 mu L of 5% hydrogen peroxide are added and uniformly mixed to obtain a chloroperoxidase catalytic system (water phase). Firstly, adding CCl with the thickness of 25mm into a container 4 (liquid phase which is mutually insoluble in water) and then to the containerSlowly adding water phase, and floating in CCl 4 On the water phase and CCl 4 Forming a liquid-liquid interface therebetween. The reaction mixture was allowed to stand in an incubator (ZSD-1090, china) at 20℃for 7 hours, during which time 5% hydrogen peroxide was added to the aqueous phase 40. Mu.L each time. The upper surface of the aqueous phase (the gas-liquid interface of the aqueous phase and air) forms a "syringic acid-3, 4-diaminobenzoic acid copolymer film", at the same time in water and CCl 4 Form a "syringic acid-3, 4-diaminobenzoic acid copolymer film".
(10) Preparation of anthocyanin-4, 4' -diaminodibenzyl copolymer membrane by chloroperoxidase catalysis:
0.020g anthocyanin and 0.010g 4,4' -diamino dibenzyl are dissolved in 100mL 0.05mol/L sodium citrate-citric acid buffer solution (pH 4.0), 1U of chloroperoxidase and 16 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain a chloroperoxidase catalytic system (water phase). Firstly, adding CCl with the thickness of 25mm into a container 4 (liquid phase which is mutually insoluble with water) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On the water phase and CCl 4 Forming a liquid-liquid interface therebetween. The reaction mixture was allowed to stand in an incubator (ZSD-1090, china) at 25℃for 8 hours, during which time 5% hydrogen peroxide was added to the aqueous phase 16. Mu.L each time. The upper surface of the aqueous phase (gas-liquid interface of the aqueous phase and air) forms an 'anthocyanin-4, 4' -diaminodibenzyl copolymer film ', and the anthocyanin-4, 4' -diaminodibenzyl copolymer film is formed on water and CCl simultaneously 4 The liquid-liquid interface of the two phases forms a "anthocyanin-4, 4' -diaminodibenzyl copolymer film".
(11) Preparation of 2, 6-dihydroxytoluene-4, 4' -binaphthyl amine copolymer film catalyzed by monophenol monooxidase:
0.050g of 2, 6-dihydroxytoluene and 0.025g of 4,4' -binaphthyl amine are dissolved in 250mL of 0.05mol/L potassium hydrogen phthalate-sodium hydroxide buffer solution (pH 5.5), 2.5U of monophenol monooxidase is added, and the mixture is uniformly mixed to obtain a monophenol monooxidase catalytic system (water phase). Firstly, adding CCl with the thickness of 25mm into a container 4 (liquid phase which is mutually insoluble with water) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On the water phase and CCl 4 Forming a liquid-liquid interface therebetween. Standing in 30 deg.C incubator (ZSD-1090, china) for reaction 1 And 0h. The upper surface of the aqueous phase (gas-liquid interface of aqueous phase and air) forms a "2, 6-dihydroxytoluene-4, 4' -binaphthyl amine copolymer film", while water, CCl 4 The liquid-liquid interface of the two phases forms a "2, 6-dihydroxytoluene-4, 4' -binaphthyl amine copolymer film".
(12) Preparation of hydroxytyrosol-guanidine copolymer membranes catalyzed by catechol oxidase:
1.000g of hydroxytyrosol and 1.250g of guanidine are dissolved in 250mL of 0.05mol/L disodium hydrogen phosphate-citric acid buffer solution (pH 5.0), and 0.005U of catechol oxidase is added and uniformly mixed to obtain a catechol oxidase catalytic system (water phase). Firstly, adding CCl with the thickness of 25mm into a container 4 (liquid phase which is mutually insoluble with water) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On the water phase and CCl 4 Forming a liquid-liquid interface therebetween. Standing in incubator (ZSD-1090, china) at 30deg.C for 30 hr. The upper surface of the aqueous phase (the gas-liquid interface of the aqueous phase and air) forms a "hydroxytyrosol-guanidine copolymer film", while water, CCl 4 The liquid-liquid interface of the two phases forms a "hydroxytyrosol-guanidine copolymer film".
(13) Preparation of 2,4, 6-trihydroxybenzaldehyde-1, 2-diaminocyclohexane copolymer membranes catalyzed by bilirubin oxidase:
0.400g of 2,4, 6-trihydroxybenzaldehyde and 0.400g of 1, 2-diaminocyclohexane are dissolved in 200mL of 0.05mol/L boric acid-borax buffer solution (pH 8.0), and bilirubin oxidase 120U is added and mixed uniformly to obtain a bilirubin oxidase catalytic system (aqueous phase). Firstly, adding CCl with the thickness of 25mm into a container 4 (liquid phase which is mutually insoluble with water) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On the water phase and CCl 4 Forming a liquid-liquid interface therebetween. Standing in oven (ZFD-5090, china) at 60deg.C for 14 hr. The upper surface of the aqueous phase (gas-liquid interface of the aqueous phase and air) forms a "2,4, 6-trihydroxybenzaldehyde-1, 2-diaminocyclohexane copolymer film", while in water, CCl 4 The liquid-liquid interface of the two phases forms a "2,4, 6-trihydroxybenzaldehyde-1, 2-diaminocyclohexane copolymer film".
Example 4: preparation of copolymer film by upper water phase-lower liquid gallium interface enzymatic polymerization
0.125g of parahydroxybenzoic acid and 0.250g of 2-fluoro-p-phenylenediamine are dissolved in 125mL of distilled water, the pH is adjusted to 6.0 by phosphoric acid, 0.125U of laccase is added, and the mixture is uniformly mixed to obtain a laccase catalytic system (water phase). A layer of liquid gallium is put into a plastic container, then a laccase catalytic system (aqueous phase) is slowly added into the container, the aqueous phase floats on the liquid gallium, and an upper aqueous phase-lower liquid gallium interface is formed between the aqueous phase and the liquid gallium. Standing in oven (ZFD-5090, china) at 40deg.C for 48 hr. The p-hydroxybenzoic acid-2-fluoro-p-phenylenediamine copolymer film is formed at the aqueous solution-liquid gallium interface.
Example 5: preparation of copolymer film by upper water phase-lower paraffin wax interface enzymatic polymerization
0.250g chlorogenic acid, 0.125g 2, 3-diaminopyridine are dissolved in 125mL of 0.05mol/L disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution (pH 7.0), and 0.125U of horseradish peroxidase and 20 mu L of 5% hydrogen peroxide are added and uniformly mixed to obtain a horseradish peroxidase catalytic system (water phase). And placing the paraffin into a container, heating to melt and level the paraffin, and standing at room temperature to cool and solidify the paraffin into a solid. To the solid paraffin in the vessel was added a horseradish peroxidase catalytic system (aqueous phase) which was located above the solidified paraffin. The reaction was carried out at 25℃with shaking at 100rpm with a constant temperature shaker (Crystal, IS-RDV3, co., ltd.) for 12 hours, during which 5% hydrogen peroxide was added to the aqueous phase several times, 20. Mu.L each. A "chlorogenic acid-2, 3-diaminopyridine copolymer film" was formed on the upper surface of the solid paraffin. At the same time, a granular "chlorogenic acid-2, 3-diaminopyridine copolymer" is produced in the aqueous phase, suspended in the aqueous phase. The aqueous phase above the membrane was decanted and the copolymer membrane was rinsed with distilled water. The vessel was placed at 70 ℃ and paraffin melted to liquid, the PE plate was inserted into the liquid paraffin, carefully moved under the "chlorogenic acid-2, 3-diaminopyridine copolymer film" and the film was gently fished out and spread flat on the PE plate.
Example 6: preparation of copolymer film by upper water phase-lower butter interface enzymatic polymerization
0.300g of tannic acid and 0.300g of ethidium bromide are dissolved in 100mL of 0.05mol/L sodium malonate-malonic acid buffer solution (pH 6.0), 0.05U of lignin peroxidase and 15 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain a lignin peroxide catalytic system (water phase). Placing beef tallow in a container, heating to melt and level, and standing at room temperature to cool and solidify the beef tallow into solid. To this vessel was added a lignin peroxide catalytic system (aqueous phase) which was located above the coagulated tallow. The reaction was carried out at 25℃with shaking at 100rpm with a constant temperature shaker (Crystal, IS-RDV3, co., ltd.) for 20 hours, during which 5% hydrogen peroxide was added to the aqueous phase several times, 15. Mu.L each time. A "tannic acid-ethidium bromide copolymer film" was formed on the upper surface of the solid tallow. At the same time, a particulate "tannic acid-ethidium bromide copolymer" is produced in the aqueous phase, suspended in the aqueous phase. The aqueous phase above the membrane was decanted and the copolymer membrane was rinsed with distilled water. The vessel was placed at 65 ℃ and the tallow melted into a liquid, the PE plate was inserted into the liquid tallow, carefully moved under the "tannic acid-ethidium bromide copolymer film" and the film was gently fished out and spread flat over the PE plate.
Example 7: preparation of copolymer film by enzymatic polymerization at water-oil phase interface
0.02g of lysine is dissolved in 50mL of 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 5.0), laccase 2U is added, and the mixture is uniformly mixed to obtain an aqueous phase part of a laccase catalytic system. 0.02g of tert-butyl hydroquinone is dissolved in 50mL of soybean oil, and is uniformly mixed to obtain an oil phase part of a laccase catalytic system, the oil phase part is covered on a water phase, and the mixture is subjected to standing reaction for 5h in a 30 ℃ incubator (Zhiyeng, ZSD-1090, china). A "terbutylhydroquinone-lysine copolymer film" formed at the interface of the aqueous phase and the oil phase.
Example 8: preparation of copolymer film by enzymatic polymerization at water-oil phase interface
0.05g of diethylenetriamine is dissolved in 50mL of 0.05mol/L trisodium phosphate-phosphoric acid buffer solution (pH 6.0), 5U of soybean peroxidase and 8 mu L of 5% hydrogen peroxide are added, and the mixture is uniformly mixed to obtain an aqueous phase part of the soybean peroxidase catalytic system. Gradually covering a layer of urushiol with a thickness of more than 1mm on the water phase, standing in an oven (Zhiyeng, ZFD-5090, china) at 40 ℃ for reaction for 5h, and adding 8 mu L of 5% hydrogen peroxide to the water phase every 1 h. A "urushiol-diethylenetriamine copolymer film" is formed at the interface of urushiol and the aqueous phase.
Example 9: preparation of multilayer films
0.040g of guaiacol and 0.025g of 4,4' -diamino dicyclohexylmethane are dissolved in 50mL of 0.05mol/L tartaric acid-sodium tartrate buffer solution (pH 7.0), and then 20U of monophenol monooxidase is added and uniformly mixed to obtain a monophenol monooxidase catalytic system (water phase). Firstly, adding CCl with the thickness of 55mm into a container 4 (liquid phase which is mutually insoluble with water), then slowly adding an aqueous phase to the vessel, the aqueous phase floating in CCl 4 And (3) upper part. Standing in 25 deg.C incubator (ZSD-1090, china) for 48 hr to form "guaiacol-4, 4' -diaminodicyclohexylmethane copolymer film" (gas-liquid interfacial film) on the upper surface of water phase (gas-liquid interface of water phase and air), and simultaneously in water phase and CCl 4 The "guaiacol-4, 4' -diaminodicyclohexylmethane copolymer film" (liquid-liquid interfacial film) was formed. The gas-liquid interface film was removed, and the solution on the upper side of the liquid-liquid interface "guaiacol-4, 4' -diaminodicyclohexylmethane copolymer film" was sucked out by a syringe, and another aqueous phase (0.125 g of gallic acid, 0.125g of 2, 4-diaminobenzenesulfonic acid, dissolved in 250mL of a 0.2mol/L disodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution (pH 7.0), and 0.5U of horseradish peroxidase and 40. Mu.L of 5% hydrogen peroxide were added and mixed well) was poured again on the upper side of the film, whereby the pipetting-pouring-pipetting operation was repeated a plurality of times. Placing in a 40 ℃ oven (zhicheng, ZFD-5090, china) again for standing reaction for 20h, adding 5% hydrogen peroxide to the water phase for multiple times, and forming a gallic acid-2, 4-diaminobenzenesulfonic acid copolymer film (gas-liquid interface film) on the upper surface of the water phase (gas-liquid interface film) each time by 40 mu L, wherein the original guaiacol-4, 4' -diaminodicyclohexylmethane copolymer film forms a gallic acid-2, 4-diaminobenzenesulfonic acid copolymer layer on the upper side of the liquid-liquid interface film, and the gallic acid-2, 4' -diaminodicyclohexylmethane copolymer film is combined with the original guaiacol-4, 4' -diaminodicyclohexylmethane copolymer film to form a double-layer film.
Example 10: enzyme-catalyzed secondary reactions (introduction of sorbitol)
Dissolving 0.1g bisphenol A, 0.1g PEI, 10mg ABTS in 70mL trisodium phosphate-phosphate buffer solution (pH 4.0) 0.05mol/L and 30mL ethanol, adding laccase0.6U, and mixing uniformly to obtain a laccase catalytic system (aqueous phase). Firstly, adding CCl with the thickness of 55mm into a container 4 (oil phase) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On the two phases, a liquid-liquid interface is formed between the two phases. Standing in 40 deg.C oven (ZFD-5090, china) for 10 hr to form bisphenol A-PEI copolymer film (gas-liquid interfacial film) on the upper surface of water phase (gas-liquid interface of water phase and air), and simultaneously mixing with water and CCl 4 The "bisphenol A-PEI copolymer film" (liquid-liquid interfacial film) is formed at the liquid-liquid interface of (C). Removing the gas-liquid interfacial film. The solution on the upper side of the "bisphenol A-PEI copolymer film" of the liquid-liquid interface was sucked out by syringe, another solution (0.3 g sorbitol was dissolved in 150mL of 0.05mol/L trisodium phosphate-phosphate buffer solution (pH 5.0), 175mg of laccase 10U, 2, 6-tetramethylpiperidine oxide (TEMPO) was added, and the mixture was homogenized), and the pipetting-pouring-pipetting operation was repeated several times. Standing in oven (ZFD-5090, china) at 40deg.C for 24 hr. Liquid-liquid (CCl) was injected with a syringe 4 ) The solution on the upper side of the interfacial film was sucked out and re-deposited in the liquid-liquid (CCl 4 ) Distilled water is injected into the upper side of the interface film, and the operations of liquid suction, liquid injection and liquid suction are repeated for a plurality of times. Inserting coverslips into CCl 4 In the middle, carefully move to liquid-liquid (CCl 4 ) Gently picking up the membrane below the interface membrane, spreading the membrane on a cover glass, and sucking residual CCl on the lower side of the membrane with filter paper 4 And (5) airing at room temperature.
The elemental composition of the film surface was measured using an X-ray photoelectron spectrometer (X-ray Photoelectron Spectroscopy, XPS, thermo Fisher Scientific, escalab 250xi, USA) and the N/O ratio of the film surface was reduced from 0.695 to 0.527 after reaction with sorbitol. The surface of the bisphenol A-PEI copolymer film which is not combined with sorbitol contains C, H, O, N elements; the sorbitol structure contains C, H, O elements, and no N element exists; thus, a decrease in the N/O ratio at the membrane surface demonstrates that the membrane incorporates sorbitol, i.e., the liquid-liquid interfacial membrane surface can undergo an enzyme-catalyzed secondary reaction.
Example 11: vesicle and enzymatic polymerization preparation method thereof
To the centrifuge tube, 100. Mu.L of oleic acid, 2mg/mL of cetyltrimethylammonium bromide (Hexadecyl trimethyl ammonium bromide, CTAB) solution (1 mL) were sequentially added, vortexed for 5min and then sonicated for 15min to prepare an oil-in-water emulsion. Centrifuging the emulsion 6000 Xg for 5min, removing the supernatant by a syringe, adding the supernatant into 1mL of 0.05mol/L trisodium phosphate-phosphoric acid buffer solution (pH 5.0) dissolved with 0.05U laccase, 2mg hydroquinone and 2mg PEI, vibrating and reacting for 24h at room temperature and in a dark place, and wrapping the hydroquinone-PEI copolymer film on the outer surface of the oil drop to obtain the hydroquinone-PEI copolymer vesicle with oleic acid wrapped inside, wherein the stability of the emulsion is greatly improved.
Soaking the prepared vesicle in ethanol, dissolving oleic acid and CTAB in the vesicle by using ethanol, centrifuging 10000 Xg for 10min, collecting the vesicle, dissolving residual oleic acid and CTAB in the ethanol again, centrifuging, and collecting the vesicle for multiple times. Evaporating the vesicle into solvent under negative pressure to obtain hollow vesicle.
Hydrophilic, hydrophobic vesicle outer walls can be prepared as desired, and target molecules/atoms can be bonded/deposited on the outer walls of the capsule.
The copolymer vesicles can be used to encapsulate cells, organelles, antigens, antibodies, functional polymers, proteins, enzymes, DNA, RNA, polysaccharides, drugs, nutrients, fragrances, pesticides, fertilizers, chemicals, fuels, explosives, and the like.
Copolymer vesicles may also provide a suitable microenvironment for some chemical or biochemical reactions.
Example 12: copolymer film reduction metal
hydroquinone-PEI vesicles were prepared as in example 11, and the vesicles were immersed in 0.05mol/L AgNO 3 Placing in a solution in a dark place for 30min, and AgNO 3 Ag in solution + The simple substance reduced into Ag nano particles is deposited on the outer surface of the vesicle wall. Centrifuging at 5000rpm for 5min, and washing with ultrapure water twice to obtain vesicles with Ag nano-particle simple substances deposited on the outer surfaces.
Those skilled in the art will appreciate that not only Ag + For other oxidizability higher than Ag + The film may also reduce and deposit it on the surface of the film,for example, noble metal ions such as gold ions, foil ions, palladium ions and the like are reduced to generate corresponding simple substances, so that the surface metallization of the electroless plating material is realized. The silver particle-covered material has excellent antibacterial performance. Palladium, platinum, silver, etc. are important chemical catalysts, and thus, the copolymer film deposited with the metal can be used for chemical catalysis. This method can also be used to remove, enrich, and recover these metals from water.
Example 13: preparation of films with conductive properties
0.25g of ferulic acid and 0.25g of p-phenylenediamine are dissolved in 125mL of 0.05mol/L trisodium phosphate-phosphoric acid buffer solution (pH 5.0), 7.5U of laccase is added, and the mixture is uniformly mixed to obtain a laccase catalytic system (water phase). Firstly, adding CCl with the thickness of 15mm into a container 4 (oil phase) then slowly add the aqueous phase to the vessel, the aqueous phase floating in the CCl 4 On water, CCl 4 A liquid-liquid interface is formed between the two phases. Standing in oven (ZFD-5090, china) at 50deg.C for 13 hr. Forming a 'ferulic acid-p-phenylenediamine copolymer film' on the upper surface of the aqueous phase (gas-liquid interface of the aqueous phase and air) while simultaneously forming a 'ferulic acid-p-phenylenediamine copolymer film' on the upper surface of the aqueous phase in water, CCl 4 The liquid-liquid interface of the two phases forms a "ferulic acid-p-phenylenediamine copolymer film". Removing the gas-liquid interfacial film. Sucking out the solution on the upper side of the liquid-liquid interface film with a syringe, and re-mixing the solution with the liquid-liquid (CCl 4 ) Distilled water is injected into the upper side of the interface film, the operations of liquid suction, liquid injection and liquid suction are repeated for a plurality of times, and a cover glass is inserted into the CCl 4 In the middle, carefully move to liquid-liquid (CCl 4 ) Gently picking up the membrane below the interface membrane, spreading the membrane on a cover glass, and sucking residual CCl on the lower side of the membrane with filter paper 4 And (5) airing at room temperature. Determination of liquid-liquid (CCl) on coverslip by four-probe method using resistance tester (lattice, ST2722, china) 4 ) The conductivity of the interfacial film surface was 1.67S/m.
The copolymer conductive film can be used in the fields of electroluminescence, electrochromic, light conduction, electronic switches, all-solid-state batteries, nonlinear optical devices, high-density memory materials, flat panel displays, organic semiconductor devices, molecular wires, light-emitting diodes, antistatic materials, electromagnetic shielding materials, photovoltaic cell materials and the like.
By selecting different film-forming monomers, copolymer films with different conductivities within a certain range can be prepared.
The conductive copolymer film can improve the conductivity of the conductive copolymer film by doping and composite deposition of metal particle simple substances.
The metal ions can be reduced on the surface of the conductive copolymer film, so that the metal simple substance is deposited on the surface of the film, and the conductivity is improved.
Comparative example 1:
(1) 0.250g bisphenol A, 0.250g pre-flavin were dissolved in 175mL of a mixed solvent of 0.1mol/L Tris-hydrochloric acid buffer solution (pH 8.5) and 75mL ethanol, and mixed well. Liquid paraffin of 5mm thickness was covered on the surface of the above solution (aqueous phase) with a pipette, and left to stand in an oven (Zhiyeng, ZFD-5090, china) at 45℃for 48 hours. No film forms at the interface between the liquid paraffin and the aqueous phase, and no particulate matter forms in the aqueous phase.
(2) 0.250g bisphenol A, 0.250g pre-flavin were dissolved in 175mL of a mixed solvent of 0.1 mol/L10 g/L NaOH solution (pH 13.4) and 75mL ethanol and mixed well. Liquid paraffin of 5mm thickness was covered on the surface of the above solution (aqueous phase) with a pipette, and left to stand in an oven (Zhiyeng, ZFD-5090, china) at 45℃for 24 hours. No film forms at the interface between the liquid paraffin and the aqueous phase, and no particulate matter forms in the aqueous phase.
Comparative example 2:
(1) 0.125g of 4,4' -dihydroxybiphenyl, 0.125g of 2, 3-diaminonaphthalene were dissolved in 215mL of a mixed solvent of 0.05mol/L glycine-sodium hydroxide buffer solution (pH 8.5) and 35mL of acetone, and mixed well. Liquid paraffin of 10mm thickness was covered on the surface of the above solution (aqueous phase) with a pipette, and left to stand in an oven (Zhiyeng, ZFD-5090, china) at 40 ℃ for 48 hours. No film forms at the interface between the liquid paraffin and the aqueous phase, and no particulate matter forms in the aqueous phase.
(2) 0.125g of 4,4' -dihydroxybiphenyl and 0.125g of 2, 3-diaminonaphthalene were dissolved in 215mL of a 10g/L NaOH solution (pH 13.4) and 35mL of acetone in a mixed solvent, and mixed well. Liquid paraffin of 10mm thickness was covered on the surface of the above solution (aqueous phase) with a pipette, and left to stand in an oven (Zhiyeng, ZFD-5090, china) at 40 ℃ for 24 hours. No film forms at the interface between the liquid paraffin and the aqueous phase, and no particulate matter forms in the aqueous phase.
The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (22)

1. A method for synthesizing a copolymer film, comprising the steps of:
dissolving a film-forming monomer containing phenolic hydroxyl groups, a film-forming monomer containing at least two amino groups and a catalyst in an aqueous phase and/or a liquid phase which is mutually insoluble in water, and polymerizing at the interface of the two phases to form a film to obtain the copolymer film;
the catalyst is at least one of peroxidase and/or oxidoreductase with laccase activity and/or artificial enzyme with catalytic activity;
The structure of the film forming monomer containing phenolic hydroxyl is shown as a formula I:
wherein each R 1 The same or different are independently selected from H, halogen, CN, NO 2 OH, SH, COOH, unsubstituted or substituted by one, two or more R a1 Substituted with the following groups: c (C) 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 Cycloalkyl, C 3-10 Cycloalkenyl, C 3-10 Cycloalkynyl radicals, C 6-14 Aryl, 5-14 membered heteroaryl, 3-10 membered heterocyclyl, -OR 1-2 、-SR 1-3 、-NR 1-4 R 1-5 、-OC(O)R 1-7 、-S(O) 2 R 1-8 、-OS(O) 2 R 1-9 、-P(O)R 1-10 R 1-11 、-N＝NR 1-12 ；
A 1 Presence or absence; when A is 1 When present, is selected from unsubstituted or substituted by one, two or more R b1 Substituted C linked to the benzoring 6-14 Aryl, 5-14 membered heteroaryl, 5-10 membered heterocyclyl;
each R a1 、R b1 The same or different, independently of one another, from H, halogen, CN, OH, SH, NO 2 、COOH、-OR 1-2 、-SR 1-3 、-NR 1-4 R 1-5 、-OC(O)R 1-7 、-S(O) 2 R 1-8 、-OS(O) 2 R 1-9 、P(O)R 1-10 R 1-11 Unsubstituted or optionally substituted by one, two or more R 1-12 Substituted C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 Cycloalkyl, C 3-10 Cycloalkenyl, C 3-10 Cycloalkynyl radicals, C 6-14 Aryl, 5-14 membered heteroaryl, 3-10 membered heterocyclyl;
each R is 1-2 、R 1-3 、R 1-4 、R 1-5 、R 1-6 、R 1-7 、R 1-8 、R 1-9 、R 1-10 、R 1-11 、R 1-12 The same or different, independently of one another, from H, halogen, CN, OH, SH, NO 2 、COOH、C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 Cycloalkyl, C 3-10 Cycloalkenyl, C 3-10 Cycloalkynyl radicals, C 6-14 Aryl, 5-14 membered heteroaryl, 3-10 membered heterocyclyl;
the film-forming monomer containing at least two amino groups has a structure shown in a formula II:
Each R 2 The same or different are independently selected from H, halogen, CN, NO 2 NO, OH, SH, COOH, unsubstituted or substituted by one, two or more R a2 Substituted with the following groups: c (C) 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 Cycloalkyl, C 3-10 Cycloalkenyl, C 3-10 Cycloalkynyl and 3-14 membered heterogeniesCyclic group, -OR 2-2 、-SR 2-3 、-NR 2-4 R 2-5 、-C(O)R 2-6 、-OC(O)R 2-7 、-S(O) 2 R 2-8 、-OS(O) 2 R 2-9 、P(O)R 2-10 R 2-11 ；
A 2 Selected from unsubstituted or optionally substituted by one, two or more R c2 Substituted C 1-6 Alkylene, ch=n-n=ch, ch=ch-CO-CH 2 -CO-CH＝CH；
p is an integer of 0 to 12;
each R a2 、R c2 Identical or different, independently of one another, from the group consisting of H, halogen, CN, OH, SH, oxo (=o), =nh, NO 2 、COOH、C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 Cycloalkyl, C 3-10 Cycloalkenyl, C 3-10 Cycloalkynyl, 3-to 10-membered heterocyclyl, -OR 2-2 、-SR 2-3 、-NR 2-4 R 2-5 、-C(O)R 2-6 、-OC(O)R 2-7 、-S(O) 2 R 2-8 、-OS(O) 2 R 2-9 、P(O)R 2-10 R 2-11 ；
Each R is 2-2 、R 2-3 、R 2-4 、R 2-5 、R 2-6 、R 2-7 、R 2-8 、R 2-9 、R 2-10 、R 2-11 The same or different, independently of one another, from H, halogen, NH 2 CN, OH, SH, oxo (=o), NO 2 、COOH、C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-10 Cycloalkyl, C 3-10 Cycloalkenyl, C 3-10 Cycloalkynyl, 3-10 membered heterocyclyl.
2. A method for synthesizing a copolymer film, comprising the steps of:
dissolving a film-forming monomer containing phenolic hydroxyl groups, a film-forming monomer containing at least two amino groups and a catalyst in an aqueous phase and/or a liquid phase which is mutually insoluble in water, and polymerizing at the interface of the two phases to form a film to obtain the copolymer film;
The catalyst is at least one of peroxidase and/or oxidoreductase with laccase activity and/or artificial enzyme with catalytic activity;
the film-forming monomer containing phenolic hydroxyl groups has a structure shown in formulas I-1 to I-8 or I-10 to I-13:
wherein R is 1 M and R c1 Having the same definition as in claim 1; b is selected from quilt R 1 Substituted N; D. d (D) 1 、D 2 And D 3 Identical or different, independently of one another, from chemical bonds, unsubstituted or optionally substituted by one or two R c1 Substituted CH 2 The method comprises the steps of carrying out a first treatment on the surface of the E and E 1 The same or different, independently of one another, are selected from the group consisting of chemical bonds, C 1-6 Alkylene, ch=ch, ch= N, N =n or ch=n-n=ch; n is an integer of 0 to 5; f (F) 1 And F 2 The same or different, independently of one another, are selected from N, CH or O + ；R 1 ’、R 1 "and R 1 Is the same as defined in the specification;
the film-forming monomer containing at least two amino groups has a structure shown in formula II-3, II-8 or II-9:
H 2 N-E 3 -NH 2
II-3、
wherein R is 2 And p has the same definition as in claim 1; q is an integer of 0 to 12; r is R 2 ' and R 2 Is the same as defined in the specification; and R is 2 、R 2 At least two of the' are amino groups;
each E 3 、E 4 Identical or different, independently of one another, from chemical bonds, unsubstituted or optionally substituted by one, two or more R c2 Substituted C 1-6 Alkylene, ch= N, CH =n-n= CH. Alkylene or alkenyl radicals having up to 6 carbon atoms-CO-C 1-6 alkylene-CO-alkylene or alkenyl having up to 6 carbon atoms, alkylene or alkenyl-CO-NH-alkylene or alkenyl having up to 6 carbon atoms, alkylene or alkenyl-NH-alkylene having up to 6 carbon atoms, alkylene or alkenyl-C (O) O-alkylene having up to 6 carbon atoms, alkylene or alkenyl having up to 6 carbon atoms.
3. A method for synthesizing a copolymer film, comprising the steps of:
dissolving a film-forming monomer containing phenolic hydroxyl groups, a film-forming monomer containing at least two amino groups and a catalyst in an aqueous phase and/or a liquid phase which is mutually insoluble in water, and polymerizing at the interface of the two phases to form a film to obtain the copolymer film;
the catalyst is at least one of peroxidase and/or oxidoreductase with laccase activity and/or artificial enzyme with catalytic activity;
the film forming monomer containing phenolic hydroxyl groups is selected from at least one of 1, 3-dihydroxynaphthalene, p-hydroxybenzoic acid, ferulic acid, 1, 6-dihydroxynaphthalene, 3-methyl salicylic acid, p-coumaric acid, 4' -dihydroxybiphenyl, bisphenol A, chlorogenic acid, 3-methoxy salicylic acid, gallic acid, tannic acid, 3, 5-diiodosalicylic acid, gentisic acid, resveratrol, esculin, p-amino salicylic acid, caffeic acid, ellagic acid, anthocyanin, syringic acid, tyrosine, guaiacol, 2, 6-dihydroxytoluene, catechol, hydroxytyrosol, galloblue, 1,8, 9-trihydroxy anthracene, urushiol and tert-butylhydroquinone;
The film forming monomer containing at least two amino groups is at least one selected from arginine, polyethyleneimine (PEI), kanamycin, diethylenetriamine, lysine, 1, 6-hexamethylenediamine, spermine, 4' -diamino dicyclohexylmethane, urea, guanidine and 1, 2-diamino cyclohexane.
4. A synthetic method according to any one of claims 1 to 3, wherein the copolymer film is obtained by dissolving a film-forming monomer containing a phenolic hydroxyl group and a film-forming monomer containing at least two amino groups in a solvent, adding a catalyst and mixing to obtain an aqueous phase, contacting the aqueous phase with a liquid phase which is insoluble in water, and polymerizing at the interface of the two phases to form a film.
5. A synthetic method according to any one of claims 1 to 3, characterized in that a film-forming monomer containing a phenolic hydroxyl group is dissolved in a solvent, a catalyst is added and mixed to obtain an aqueous phase, the film-forming monomer containing at least two amino groups is dissolved in a liquid phase which is mutually insoluble in water, the aqueous phase is contacted with the liquid phase which is mutually insoluble in water, and the copolymer film is polymerized at the interface of the two phases.
6. A synthetic method according to any one of claims 1 to 3, characterized in that a film-forming monomer containing at least two amino groups is dissolved in a solvent, a catalyst is added and mixed to obtain an aqueous phase, a film-forming monomer containing a phenolic hydroxyl group is dissolved in a liquid phase which is mutually insoluble in water, the aqueous phase is contacted with a liquid phase which is mutually insoluble in water, and the copolymer film is polymerized at the interface of the two phases.
7. A synthetic method according to any one of claims 1 to 3, characterized in that a catalyst is dissolved in an aqueous phase, a film-forming monomer containing a phenolic hydroxyl group, a film-forming monomer containing at least two amino groups are dissolved in a liquid phase which is mutually insoluble in water, the aqueous phase is brought into contact with the liquid phase which is mutually insoluble in water to undergo an enzymatic reaction, and the copolymer film is polymerized at the interface of the two phases to form a film.
8. The synthetic method according to any one of claims 1 to 7, wherein the peroxidase is selected from at least one of manganese peroxidase, lignin peroxidase, chloroperoxidase and plant peroxidase; the plant peroxidase is at least one of horseradish peroxidase, soybean peroxidase, rice peroxidase, cotton peroxidase, safflower bean peroxidase, chickpea peroxidase, guar peroxidase and pea peroxidase;
the oxidoreductase with laccase activity is selected from at least one of catechol oxidase, monophenol monooxidase and bilirubin oxidase;
the artificial enzyme is an enzyme mimic with specific catalytic function synthesized by utilizing an organic chemistry and biology method according to the catalytic mechanism of the enzyme and simulating the biological catalytic function of natural enzyme.
9. The method of claim 8, wherein the laccase is a fungal laccase.
10. The method of claim 9, wherein the fungal laccase is a polyporus winter laccase.
11. The method of claim 1, wherein when the catalyst is at least one of laccase, bilirubin oxidase, and peroxidase, the combination of the phenolic hydroxyl group-containing film-forming monomer and the film-forming monomer containing at least two amino groups is: r is R 1 And/or A 1 A combination of a compound of formula I and a compound of formula II when there is at least one phenolic hydroxyl group in the structure;
when the catalyst is a monophenol monooxidase, the combination of the film-forming monomer containing a phenolic hydroxyl group and the film-forming monomer containing two amino groups is: a combination of a compound of formula I and a compound of formula II;
when the catalyst is catechol oxidase, the combination of the film-forming monomer containing a phenolic hydroxyl group and the film-forming monomer containing at least two amino groups is: r is R 1 A combination of a compound of formula I and a compound of formula II when the compound is a phenolic hydroxyl group in an ortho position;
the film-forming monomer containing phenolic hydroxyl and the film-forming monomer containing at least two amino groups are respectively one or more than two in a reaction system.
12. A method according to any one of claims 1 to 3, wherein the aqueous phase is an aqueous solution having a water content of not less than 50% by total mass.
13. The method of claim 12, wherein the aqueous phase is dissolved with a film-forming monomer and contains a water-soluble organic solvent, the organic solvent being at least one of methanol, ethanol, isopropanol, acetone, methyl formate, ethyl acetate, acetonitrile, tetrahydrofuran, N-dimethylformamide, 1, 4-dioxane, dimethyl sulfoxide, diethylene glycol butyl ether, diethylene glycol.
14. The method of claim 7, wherein the enzymatic reaction further incorporates at least one of an enhancer for the enzyme, hydrogen peroxide, metal ions, buffer ion pairs.
15. The method of claim 7, wherein the water-immiscible liquid phase is selected from the group consisting of an oil that is a liquid at ambient temperature, an oil that is a solid at a set temperature, and a liquid metal.
16. The method of claim 4, wherein the solvent is selected from the group consisting of water and buffer solutions, wherein the buffer solution is a sodium acetate-acetic acid buffer solution, a disodium hydrogen phosphate-citric acid buffer solution, a potassium hydrogen phthalate-sodium hydroxide buffer solution, a tartaric acid-sodium tartrate buffer solution, a sodium citrate-citric acid buffer solution, a trisodium phosphate-phosphoric acid buffer solution, a sodium malonate-malonic acid buffer solution, a sodium succinate-succinic acid buffer solution, a phthalic acid-hydrochloric acid buffer solution, a disodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution, a disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution, a dipotassium hydrogen phosphate-sodium hydroxide buffer solution, a Tris-hydrochloric acid buffer solution, a boric acid-borax buffer solution, and a glycine-sodium hydroxide buffer solution.
17. A process according to any one of claims 1 to 3, wherein the catalyst is used in the reaction system in an amount of from 0.01 to 600U/L in terms of enzyme activity.
18. A process according to any one of claims 1 to 3, wherein the catalyst is used in the reaction system in an amount of from 0.5U/L to 200U/L in terms of enzyme activity.
19. A method according to any one of claims 1 to 3, wherein the film-forming monomer is present in the reaction system at a mass concentration of 0.002 to 380g/L.
20. A method according to any one of claims 1 to 3, wherein the temperature of the film forming reaction is 4 to 90 ℃.
21. A method according to any one of claims 1 to 3, wherein the film forming reaction is carried out for a period of 0.01 to 72 hours.
22. A process according to any one of claims 1 to 3, wherein the pH of the aqueous phase is from 2 to 10.
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