CN109439189A - A kind of process for fixation of antifouling enzyme on inorganic coating surface - Google Patents
A kind of process for fixation of antifouling enzyme on inorganic coating surface Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
- C09D183/00—Coating compositions based on macromolecular compounds obtained by reactions forming in the main chain of the macromolecule a linkage containing silicon, with or without sulfur, nitrogen, oxygen, or carbon only; Coating compositions based on derivatives of such polymers
- C09D183/04—Polysiloxanes
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
- C09D4/00—Coating compositions, e.g. paints, varnishes or lacquers, based on organic non-macromolecular compounds having at least one polymerisable carbon-to-carbon unsaturated bond ; Coating compositions, based on monomers of macromolecular compounds of groups C09D183/00 - C09D183/16
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
- C09D5/00—Coating compositions, e.g. paints, varnishes or lacquers, characterised by their physical nature or the effects produced; Filling pastes
- C09D5/08—Anti-corrosive paints
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
- C09D5/00—Coating compositions, e.g. paints, varnishes or lacquers, characterised by their physical nature or the effects produced; Filling pastes
- C09D5/16—Antifouling paints; Underwater paints
- C09D5/1606—Antifouling paints; Underwater paints characterised by the anti-fouling agent
- C09D5/1612—Non-macromolecular compounds
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
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Abstract
The invention belongs to antifouling technical field of enzyme immobilization, a kind of process for fixation more particularly to antifouling enzyme on inorganic coating surface, use alkoxy silane, amino silane, contain the inorganic coating of amino group in silica solution preparation surface, antifouling enzyme is fixed on transparent silica solution inorganic coating surface by the connection function of glutaraldehyde, inorganic coating is set to have preferable anti-pollution while with good antiseptic property, applied to marine ship, works (dock, drilling platforms, pier and stake) etc. surface soiling and the key positions such as underwater kit observation window anticorrosion and antifouling, technical support can also be provided for the preparation of enzyme base nonpolluting coating material；Preparation method simple possible, it is raw materials used cheap and easily-available, cost of manufacture is greatly reduced, the antifouling enzyme of silica solution inorganic coating immobilization has the preferable transparency and good anti-corrosion and anti-pollution, has a extensive future in terms of the surface soiling and anticorrosion and antifouling in marine environment.
Description
Technical field:
The invention belongs to antifouling technical field of enzyme immobilization, and in particular to a kind of fixation of antifouling enzyme on inorganic coating surface
Change method is contained the inorganic coating of amino group using alkoxy silane, amino silane, silica solution preparation surface, is based on penta 2
Antifouling enzyme is formed antifouling enzyme and is covalently fixed on inorganic coating surface by the connection function of aldehyde.
Background technique:
Marine fouling organism is also referred to as marine attaching organism, be grow alow with the animal on all marine facility surfaces,
The general name of plant and microorganism, fouling organism apposition growth process are known as biodeterioration.Marine biofouling is to ship, underwater dress
Standby and marine Artificial facilities cause to seriously endanger, it is necessary to which takeing effective measures prevents marine organisms to be stained attachment caused by it.
Organotin antifouling paints have been prohibited from using because seriously endangering marine ecology and environment；Widely used cupric in the prior art
Element antifouling paint also results in serious marine eco-environment problem due to will lead to the accumulation of bay copper, in some states
Family is used by limitation and faces the trend of disabling.Ship and marine Artificial facilities are caused seriously to endanger for marine biofouling
The background that harmful and conventional toxic antifouling paint faces disabling and limitation uses, developing environmentally friendly marine antifouling coating has
Significance, wherein enzyme base nonpolluting coating technology has caused countries in the world to be closed extensively due to its feature of environmental protection, high efficiency and universal
Note and further investigation become one of the important directions of novel environmentally friendly anti-fouling material development.The study found that marime fouling is raw
Object attached base is more or less the same, and adhesion substances include the substances such as protein-based, polysaccharide and lipid.Decomposition based on enzyme is made
The effective inhibiting effect of generation can be adhered to it with the direct adhesion substances that decompose, such as proteases for decomposing protide sticks object
Matter, Amylase Hydrolysis polysaccharide adhesion substances, lipase decompose lipid adhesion substances etc..Object is sticked to fouling organism using enzyme
The efficient hydrolysing activity of matter and good environmental-friendly sexual development enzyme base nonpolluting coating material are marine anti-pollution technology important developments
Trend, antifouling enzyme immobilizatio processing are the important links in its application process.
The common fixing means of antifouling enzyme is divided into physisorphtion, investment, cross-linking method and covalent knot according to active force
It is legal.Enzyme immobilization carrier mainly has an inorganic carrier and organic carrier two major classes, and inorganic carrier mainly has diatomite, silica gel, more
The porous inorganic materials such as hole glass and mesopore molecular sieve, organic carrier mainly include natural polymer subcarrier (such as sodium alginate, fibre
Tie up element, chitosan etc.) and synthetic organic polymer carrier (such as macroreticular resin, synthetic fibers).The confirmations such as Gatenholm embedding
Bacterium in coating can still maintain vigour and generate bioactive ingredients；Holmstrom etc. then inquired into influence be fixed on it is solidifying
The factor of bacterial action and required condition in glue；Lipase from candida sp is covalently fixed on silicon respectively by Dandavate, Yilmaz
Nanoparticle and glass microballoon surface obtain reusable and for lactate synthesis enzyme preparation；Huijs et al. passes through polymerization
Acetoacetyl, aldehyde radical, chloromethyl in object etc. carry out enzyme immobilizatio.Immobilised enzymes and antifouling test result show to pass through
The functional group of adhesive coating, which is covalently attached immobilized enzyme, can generate effective antifouling effect, and the enzyme applied in this way is applying
Significant anti-fouling effect is embodied in layer, in addition immobilization also improves enzyme anti-pollution.The U.S. of Biolocus company application
Patent US20080038241 disclose it is a kind of use organosilicon-acrylic acid from polishing resin as film forming matter, generated with enzymatic
Oxide makees the antifouling paint of anti-fouling agent；South Korea's university research makees anti-dirt with business fund application enzyme and fibre blend
The antifouling paint of matter；GENECOR company, internationally recognizable Denmark's enzyme preparation company and famous HEMPEL company, coating material production manufacturer
Antifouling new technology of the R & D Cooperation based on glucase and hexoxidase synergy production hydrogen peroxide, the technology has been at present
Through being successfully applied in antifouling paint, and real ship application is started.
The anticorrosive paint of marine anticorrosion field application mainly has epoxies, polyurethanes, rubber, fluorine type resin, organic
Silicones class, polyurea elastomer class etc., under water in marine environment, the application usually in a manner of matching system, such as anti-corrosion bottom
Paint, intermediate connection paint, antifouling topcoat system, i.e. corrosion-inhibiting coating and nonpolluting coating pass through connection paint bonding, mating application.It is Chinese special
For a kind of antifouling enzyme disclosed in benefit 201810373368.0 in the process for fixation of surface of graphene oxide, technical process includes point
It dissipates, totally four steps, critical process are antifouling enzyme consolidating on graphene oxide powder surface for amination, aldehyde radical and immobilization
Fixedization is used to prepare anti-sea organism attachment material, does not provide Corrosion Protection；Disclosed in Chinese patent 201610250733.X
It is a kind of covalently to fix the temperature sensitive from polishing process for preparing resins of antifouling enzyme, including temperature sensitive from the preparation for polishing resin and covalently fixed
Two steps of antifouling enzyme are fixed wherein covalently fixing antifouling enzyme process and being related to antifouling enzyme in resin particle surface, contain enzyme after fixed
Resin is scattered in xylene solution again, forms anti-sea organism attachment coating resin, this resin can be by under marine environment
One-step hydrolysis only has single anti-pollution function, does not provide Corrosion Protection, in addition the transparency of coating is poor, does not also have
The antifouling application potential of underwater viewing window.The above patented method is not suitable for antifouling enzyme in the immobilization of organic coating layer surface, such as
A kind of coating of fruit is provided simultaneously with good antiseptic property and anti-pollution, will significantly reduce cost and expand application
Field, the application of this aspect is still rarely reported at present.Therefore, according to marine ship, marine structure, underwater kit observation window etc.
Urgent need of the marime fouling environment to green nonpolluting coating proposes a kind of antifouling enzyme in the immobilization side on inorganic coating surface
Method.
Summary of the invention:
It is an object of the invention to overcome disadvantage of the existing technology, a kind of antifouling enzyme of R & D design is in inorganic coating table
The process for fixation in face so that the inorganic coating covalently fixed after antifouling enzyme also have while having good antiseptic property compared with
Good anti-pollution.
To achieve the goals above, the technique mistake of process for fixation of the antifouling enzyme of the present invention on inorganic coating surface
Journey includes preparing organic coating, crosslinking Treatment and the antifouling enzyme of immobilization totally three steps:
(1) it prepares organic coating: alkoxy silane being added in the silica solution tune that pH value is 4.0-5.0, in 45-60 DEG C
Lower hydrolysis 0.5-2h forms mixed liquor, aminoalkoxysilane is added in mixed liquor, is coated in pellicula after mixing
On, organic coating is formed on pellicula；
The mass values of silica solution and alkoxy silane are 0.8-1.3, and the quality of aminoalkoxysilane accounts for mixed liquor
The 0.1-5.0% of quality；
(2) crosslinking Treatment: glutaraldehyde is added in the phosphate buffer that pH value is 7, it is dense is configured to mass percent
The glutaraldehyde solution that degree is 5%, pellicula is immersed in glutaraldehyde solution, is carried out the glutaraldehyde cross-linking processing of 5h, is then used
The phosphate buffer that pH value is 7 cleans pellicula 3 times, removes remaining glutaraldehyde, completes crosslinking Treatment；
(3) the antifouling enzyme of immobilization: antifouling enzyme is added in the phosphate buffer that pH value is 7, is configured to quality percentage
Specific concentration is the antifouling enzyme solutions of 5-20%, the pellicula after step (2) crosslinking Treatment is immersed in antifouling enzyme solutions, in 5-
5h is impregnated at 30 DEG C, is then cleaned pellicula 3 times with the phosphate buffer that pH value is 7, the sample after cleaning is placed in vacuum
It is freeze-dried in freeze drier, completes antifouling enzyme in the immobilization of organic coating layer surface.
Alkoxy silane of the present invention is ethyl orthosilicate alkane, methyltriethoxysilane and methyl trimethoxy oxygroup silicon
One or more of alkane.
Aminoalkoxysilane of the present invention is diethylenetriamine base propyl trimethoxy silicane, (3- aminopropyl) three
One or more of methoxy silane and (3- aminopropyl) triethoxysilane.
Antifouling enzyme of the present invention is one or more of protease, amylase, chitinase and lysozyme.
Compared with prior art, the present invention containing amino base using alkoxy silane, amino silane, silica solution preparation surface
The inorganic coating of group, is fixed on transparent silica solution inorganic coating surface by the connection function of glutaraldehyde for antifouling enzyme, makes nothing
Organic coating has preferable anti-pollution while with good antiseptic property, is applied to marine ship, works (ship
Depressed place, drilling platforms, pier and stake) etc. surface soiling and the key positions such as underwater kit observation window anticorrosion and antifouling, additionally it is possible to
Preparation for enzyme base nonpolluting coating material provides technical support；Preparation method simple possible, it is raw materials used cheap and easily-available, significantly
It reduces the production cost, the antifouling enzyme of silica solution inorganic coating immobilization has the preferable transparency and good anti-corrosion and soil resistance
Can, it has a extensive future in terms of the surface soiling and anticorrosion and antifouling in marine environment.
Detailed description of the invention:
Fig. 1 is process flow diagram of the invention.
Specific embodiment:
The present invention is described further by way of example and in conjunction with the accompanying drawings.
Embodiment 1:
The antifouling enzyme that the present embodiment is related to is organic including preparing in the technical process of the process for fixation on inorganic coating surface
Coating, crosslinking Treatment and the antifouling enzyme of immobilization totally three steps:
(1) it prepares organic coating: ethyl orthosilicate alkane being added in the silica solution tune that pH value is 4.0, hydrolyzed at 45 DEG C
1h forms mixed liquor, and diethylenetriamine base propyl trimethoxy silicane is added in mixed liquor, is coated in film after mixing
On piece forms organic coating on pellicula；
The mass values of silica solution and ethyl orthosilicate alkane are 0.8, the matter of diethylenetriamine base propyl trimethoxy silicane
Amount accounts for the 0.1% of the quality of mixed liquor；
(2) crosslinking Treatment: glutaraldehyde is added in the phosphate buffer that pH value is 7, it is dense is configured to mass percent
The glutaraldehyde solution that degree is 5%, pellicula is immersed in glutaraldehyde solution, is carried out the glutaraldehyde cross-linking processing of 5h, is then used
The phosphate buffer that pH value is 7 cleans pellicula 3 times, removes remaining glutaraldehyde, completes crosslinking Treatment；
(3) the antifouling enzyme of immobilization: protease is added in the phosphate buffer that pH value is 7, is configured to quality percentage
The protein enzyme solution that specific concentration is 20% immerses the pellicula after step (2) crosslinking Treatment in protein enzyme solution, at 5 DEG C
5h is impregnated, is then cleaned pellicula 3 times with the phosphate buffer that pH value is 7, the sample after cleaning is placed in vacuum refrigeration and is done
It is freeze-dried in dry machine, completes antifouling enzyme in the immobilization of organic coating layer surface.
Embodiment 2:
The antifouling enzyme that the present embodiment is related to is organic including preparing in the technical process of the process for fixation on inorganic coating surface
Coating, crosslinking Treatment and the antifouling enzyme of immobilization totally three steps:
(1) it prepares organic coating: methyltriethoxysilane being added in the silica solution tune that pH value is 5.0, at 50 DEG C
2h is hydrolyzed, mixed liquor is formed, (3- aminopropyl) trimethoxy silane is added in mixed liquor, is coated in pellicula after mixing
On, organic coating is formed on pellicula；
Silica solution and the mass values of methyltriethoxysilane are 1.3, the quality of (3- aminopropyl) trimethoxy silane
Account for the 3.0% of the quality of mixed liquor；
(2) crosslinking Treatment: glutaraldehyde is added in the phosphate buffer that pH value is 7, it is dense is configured to mass percent
The glutaraldehyde solution that degree is 5%, pellicula is immersed in glutaraldehyde solution, is carried out the glutaraldehyde cross-linking processing of 5h, is then used
The phosphate buffer that pH value is 7 cleans pellicula 3 times, removes remaining glutaraldehyde, completes crosslinking Treatment；
(3) the antifouling enzyme of immobilization: protease is added in the phosphate buffer that pH value is 7, is configured to quality percentage
The protein enzyme solution that specific concentration is 10% immerses the pellicula after step (2) crosslinking Treatment in protein enzyme solution, at 10 DEG C
Then lower immersion 5h is cleaned pellicula 3 times with the phosphate buffer that pH value is 7, the sample after cleaning is placed in vacuum refrigeration
It is freeze-dried in drying machine, completes antifouling enzyme in the immobilization of organic coating layer surface.
Embodiment 3:
The antifouling enzyme that the present embodiment is related to is organic including preparing in the technical process of the process for fixation on inorganic coating surface
Coating, crosslinking Treatment and the antifouling enzyme of immobilization totally three steps:
(1) it prepares organic coating: methyltrimethoxysilane being added in the silica solution tune that pH value is 5.0, at 60 DEG C
0.5h is hydrolyzed, mixed liquor is formed, (3- aminopropyl) triethoxysilane is added in mixed liquor, is coated in applies after mixing
On diaphragm, organic coating is formed on pellicula；
Silica solution and the mass values of methyltrimethoxysilane are 1.0, the matter of (3- aminopropyl) triethoxysilane
Amount accounts for the 5.0% of the quality of mixed liquor；
(2) crosslinking Treatment: glutaraldehyde is added in the phosphate buffer that pH value is 7, it is dense is configured to mass percent
The glutaraldehyde solution that degree is 5%, pellicula is immersed in glutaraldehyde solution, is carried out the glutaraldehyde cross-linking processing of 5h, is then used
The phosphate buffer that pH value is 7 cleans pellicula 3 times, removes remaining glutaraldehyde, completes crosslinking Treatment；
(3) the antifouling enzyme of immobilization: protease is added in the phosphate buffer that pH value is 7, is configured to quality percentage
The protein enzyme solution that specific concentration is 5% immerses the pellicula after step (2) crosslinking Treatment in protein enzyme solution, at 30 DEG C
5h is impregnated, is then cleaned pellicula 3 times with the phosphate buffer that pH value is 7, the sample after cleaning is placed in vacuum refrigeration and is done
It is freeze-dried in dry machine, completes antifouling enzyme in the immobilization of organic coating layer surface.The antifouling enzyme of immobilization manufactured in the present embodiment is logical
Cross neutral salt spray method carry out corrosion resisting property test the result is that without significant corrosion in 1000h.
When the antifouling enzyme of immobilization manufactured in the present embodiment inhibit the test of diatom adhesion property, to be free of the load of protease
Slide is control blank, and the concentration of benthic diatom is diluted to 1.0 × 10 with sterilizing seawater6A/ml, diatom is right after 2 hours
Adhesion amount according to blank slide surface is 144/mm2, diatom is 31 in the adhesion amount of the antifouling enzyme slide surface of immobilization
A/cm2, test result shows that immobilization proteinase has good inhibiting effect to the attachment of diatom.
Claims (4)
1. a kind of antifouling enzyme is in the process for fixation on inorganic coating surface, it is characterised in that technical process includes preparing organic painting
Layer, crosslinking Treatment and the antifouling enzyme of immobilization totally three steps:
(1) it prepares organic coating: alkoxy silane being added in the silica solution tune that pH value is 4.0-5.0, be lauched in 45-60 DEG C
0.5-2h is solved, mixed liquor is formed, aminoalkoxysilane is added in mixed liquor, is coated on pellicula after mixing, is applied
Organic coating is formed on diaphragm；
The mass values of silica solution and alkoxy silane are 0.8-1.3, and the quality of aminoalkoxysilane accounts for the quality of mixed liquor
0.1-5.0%；
(2) crosslinking Treatment: glutaraldehyde is added in the phosphate buffer that pH value is 7, being configured to mass percent concentration is
5% glutaraldehyde solution, pellicula is immersed in glutaraldehyde solution, is carried out the glutaraldehyde cross-linking processing of 5h, is then used pH value
It is cleaned pellicula 3 times for 7 phosphate buffer, removes remaining glutaraldehyde, complete crosslinking Treatment；
(3) the antifouling enzyme of immobilization: antifouling enzyme is added in the phosphate buffer that pH value is 7, it is dense is configured to mass percent
Degree is the antifouling enzyme solutions of 5-20%, the pellicula after step (2) crosslinking Treatment is immersed in antifouling enzyme solutions, at 5-30 DEG C
Then lower immersion 5h is cleaned pellicula 3 times with the phosphate buffer that pH value is 7, the sample after cleaning is placed in vacuum refrigeration
It is freeze-dried in drying machine, completes antifouling enzyme in the immobilization of organic coating layer surface.
2. antifouling enzyme according to claim 1 is in the process for fixation on inorganic coating surface, it is characterised in that the alcoxyl
Base silane is one or more of ethyl orthosilicate alkane, methyltriethoxysilane and methyltrimethoxysilane.
3. antifouling enzyme according to claim 1 is in the process for fixation on inorganic coating surface, it is characterised in that the amino
Alkoxy silane is diethylenetriamine base propyl trimethoxy silicane, (3- aminopropyl) trimethoxy silane and (3- aminopropyl)
One or more of triethoxysilane.
4. antifouling enzyme according to claim 1 is in the process for fixation on inorganic coating surface, it is characterised in that described antifouling
Enzyme is one or more of protease, amylase, chitinase and lysozyme.
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| CN113952792A (en) * | 2020-07-01 | 2022-01-21 | 山东中昊控股集团有限公司 | Material for negative pressure purification and toxin filtration and preparation method thereof |
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|---|---|---|---|---|
| EP3878280A1 (en) * | 2020-03-12 | 2021-09-15 | Shenzhen Orient Technology Co., Ltd | Antibacterial substrate and preparation method thereof |
| CN113952792A (en) * | 2020-07-01 | 2022-01-21 | 山东中昊控股集团有限公司 | Material for negative pressure purification and toxin filtration and preparation method thereof |
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| CN109439189B (en) | 2020-12-18 |
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