CN109517090A - A kind of preparation method of the degree of polymerization and the controllable labyrinth chitosan oligosaccharide of deacetylation - Google Patents
A kind of preparation method of the degree of polymerization and the controllable labyrinth chitosan oligosaccharide of deacetylation Download PDFInfo
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
- C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
- C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
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- C07H5/00—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium
- C07H5/04—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium to nitrogen
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Abstract
The present invention relates to technical field of fine, and in particular to the preparation method of a kind of degree of polymerization and the controllable labyrinth chitosan oligosaccharide of deacetylation.Chitin is set to dissolve in certain solvent or sufficiently be swollen first, and even phase degrades to obtain small molecule chitin or chitin oligo saccharide under acid catalysis.Small molecule chitin or chitin oligo saccharide further regulate and control its degree of polymerization and deacetylation using chitinase, chitosan enzyme, chitin deacetylase or non-specific commercial enzyme containing above-mentioned enzymolysis activity, it is spray-dried to obtain the degree of polymerization 20 hereinafter, deacetylation regulatable labyrinth chitosan oligosaccharide between 20~80%.Preparation method of the present invention, which can be realized, regulates and controls the sugar chain molecules structure such as the degree of polymerization of complicated chitosan oligosaccharide and deacetylation, and product yield is high, and by-product is few, at low cost, is easy to mass production.
Description
Technical field
The present invention relates to technical field of fine, and in particular to a kind of degree of polymerization and the controllable labyrinth of deacetylation
The preparation method of chitosan oligosaccharide.
Background technique
Chitin (Chitin) is also known as chitin, and by number, equal N-acetylglucosamine (GlcNAc) does not pass through β -1,
High molecular polymer made of 4- glucosides key connection, it is close with the structure of cellulose, be widely present Yu Haiyang crustacean shell,
In insect wing, mushroom and alga cells wall, be tellurian reserves be only second to cellulose natural polysaccharide and it is second largest can
Regenerated resources.According to reckoning, the biology of nature can produce 100,000,000,000 tons of chitins every year, wherein being utilized only by modernization industry
Up to 2,000,000 tons.With the exhaustion of fossil resource and the continuous improvement of cost, developed using biomass resources such as chitins as raw material
Fine chemicals has important research and application value.
Chitosan is de-acetyl chitin product, is only a large amount of existing positively charged basic amine group polysaccharide, raw
Object compatibility is good, has many special physics, chemistry and physiological activity.The relative molecular mass of chitosan is usually in hundreds of thousands
It is extremely up to a million, and molecular structure is close, cannot be dissolved in some usual vehicles such as water, not degradable absorption in vivo, to influence
Its application range.
Chitosan oligosaccharide is by 2~20 Glucosamines, N-acetylglucosamine or combination with β -1,4 glucosides
Oligomer made of key connection.Compared with chitosan, chitosan oligosaccharide good water solubility is easily absorbed by the body, and there are many important lifes
Object activity, with the superiority that chitosan is incomparable.Chitosan oligosaccharide can be directly absorbed by animal intestinal tract epithelial cell, human body
Absorptivity can reach 99.88%, shows antibacterial, antitumor, Adjust-blood lipid, adjusts immune and activation intestinal bifidobacteria etc. and is a variety of
Physiological function.Chitosan oligosaccharide can stimulate the immune system response of plant, activate defense reaction, generate the active matter with disease resistance
Matter inhibits the formation of disease, induces the disease resistance of plant, generates immune and killing effect to a variety of fungies, bacterium and virus.Make
For a kind of natural plants inducer, chitosan oligosaccharide can enhance plant to the defence capability of pest and disease damage.After handling crops or its seed,
Seed can be excited to do sth. in advance rudiment, promote plant growth, increase the resistance of plant, improve yield.Chitosan oligosaccharide can be widely applied to
All various aspects such as plant growth regulating, feed addictive, seed treatment, agricultural products fresh-keeping, disease and insect resistance, and have it is nontoxic,
It is harmless, pollution-free can safe disposal the features such as.Chitosan oligosaccharide product is in chemistry, medicine, food, cosmetics, agricultural, ring at present
All various aspects such as guarantor, poultry industry are applied.
Now chitosan oligosaccharide in the market is mainly as Glucosamine through completely deacetylated made of β -1,4 glucosides key connection
Product.The amino of numerous studies discovery, chitosan oligosaccharide is (multiple by the chitosan oligosaccharide after complete acetylation (chitin oligo saccharide) or partial acetylation
Miscellaneous structure chitosan oligosaccharide) often there is stronger biological activity, and the chitosan oligosaccharide activity difference of different polymerization degree and deacetylation
The effect of larger or even some active functions is opposite.Such as: it has been found that chitosan oligosaccharide antibacterial activity, to the growth inhibition of tumour cell
Effect etc. and the degree of polymerization and deacetylation etc. are closely related.The complete deacetylated chitosan oligosaccharide that the degree of polymerization is 2~8, which has, inhibits tumour blood
The ability that pipe is formed, and optimal inhibition activity is then shown when its deacetylation is reduced to 70%.This structure of system research
The chitosan oligosaccharide structure-activity relationship and its molecular mechanism of action of differentiation are to develop the core place of novel chitosan oligosaccharide product.It is complicated at present
Structural shell oligosaccharide product has become the new trend of chitin industrial research in the world and market exploitation.
Prepare labyrinth chitosan oligosaccharide key be to the sugar chain molecules such as the degree of polymerization of chitosan oligosaccharide and deacetylation structure into
Row regulation.Currently, the preparation approach of chitosan oligosaccharide be mainly chitosan acid degradation (patent document such as: JPS6121103,
KR100296738), hydrogen peroxide oxidation degradation (patent document such as: CN200410020831.1, CN201010172203.0,
CN201110125040.5) and enzymatic hydrolysis (patent document such as: US4970150, CN200510019438.5, CN01126457.8,
CN201010172203.0, CN200710051931.4) etc. technology paths.Chitin produces chitosan through highly basic high temperature is deacetylated
Process be typical heterogeneous deacetylation, the acetyl group distributing inhomogeneity on chitosan chain.Degradation technologies of chitosan,
Such as: acid degradation method simple process, redox edman degradation Edman are not necessarily to strong acid and strong base, and enzyme edman degradation Edman reaction condition is mild, without side reaction
Generation, environmental pollution are small.But these reactions are all to carry out in a random basis, it is difficult to realize effective control to reaction process, cause
Keep product degree of polymerization distribution wide and Effective Regulation can not be carried out to deacetylation.Due to cannot achieve the chitosan oligosaccharide degree of polymerization and taking off
Effective control of acetyl degree etc., the prior art are difficult to meet the system for labyrinth chitosan oligosaccharide bioactivity and deeply grind
Study carefully and industrialization production demand.
Summary of the invention
The object of the present invention is to provide a kind of degree of polymerization and the preparation method of the controllable labyrinth chitosan oligosaccharide of deacetylation,
Solve it is existing in the prior art be difficult to realize the chitosan oligosaccharide degree of polymerization and deacetylation etc. effectively control the problems such as.
Present invention provide the technical scheme that
The preparation method of a kind of degree of polymerization and the controllable labyrinth chitosan oligosaccharide of deacetylation, makes chitin in solvent first
Middle dissolution or sufficiently swelling, even phase degrades to obtain small molecule chitin or chitin oligo saccharide under acid catalysis, small molecule chitin or
Chitin oligo saccharide chitinase, chitosan enzyme, chitin deacetylase or the non-specific commodity containing above-mentioned enzymolysis activity
Enzyme further regulates and controls the degree of polymerization and deacetylation, it is spray-dried obtain the degree of polymerization 20 hereinafter, deacetylation 20~
Regulatable labyrinth chitosan oligosaccharide between 80%.
The preparation method of the controllable labyrinth chitosan oligosaccharide of the degree of polymerization and deacetylation, the specific steps are as follows:
1) it is scattered in after chitin is crushed with pulverizer in the reaction dissolvent of 5 times to 50 times quality, under strong stirring, first
Shell element is sufficiently swollen or dissolves in reaction dissolvent；
2) reactant keeps the temperature 1 to 10 hour at 0 DEG C~100 DEG C, and chitin even phase degradation under acid catalysis, the degree of polymerization is with anti-
It is gradually reduced between seasonable, until obtaining small molecule chitin or chitin oligo saccharide solution；
3) reaction dissolvent is removed using the evaporation of vacuum decompression solvent, alcohol precipitation and vacuum drying, spray drying or forced air drying,
And it is dried to obtain small molecule chitin or chitin oligo saccharide；
4) small molecule chitin or chitin oligo saccharide chitinase, chitosan enzyme, chitin deacetylase or containing upper
The non-specific commercial enzyme for stating enzymolysis activity further regulates and controls the degree of polymerization and deacetylation of oligosaccharides；It is spray-dried to handle to obtain the final product
To the degree of polymerization 20 hereinafter, deacetylation regulatable labyrinth chitosan oligosaccharide between 20~80%.
The preparation method of the controllable labyrinth chitosan oligosaccharide of the degree of polymerization and deacetylation, chitin are dried through rinsing
Afterwards, then using pulverizer 80~200 mesh are crushed to.
The preparation method of the controllable labyrinth chitosan oligosaccharide of the degree of polymerization and deacetylation, dissolves the solvent of chitin
For hexafluoroisopropanol, Hexafluoro acetone, dichlroroethanol, the dimethylformamide containing lithium chloride, the dimethyl second containing lithium chloride
Amide, N- methyl morpholine oxide, concentrated hydrochloric acid, the concentrated sulfuric acid, concentrated phosphoric acid, methane sulfonic acid, formic acid, dichloroacetic acid, in trichloroacetic acid
One or more kinds of mixtures, solvent usage are 5~50 times of chitin quality.
The preparation method of the controllable labyrinth chitosan oligosaccharide of the degree of polymerization and deacetylation, when molten using faintly acid
Agent: hexafluoroisopropanol, Hexafluoro acetone, dichlroroethanol, the dimethylformamide containing lithium chloride, the dimethyl second containing lithium chloride
Amide, N- methyl morpholine oxide, formic acid, dichloroacetic acid, trichloroacetic acid one or more when, pass through and hydrochloric acid or sulphur be added
Acid, to improve the degradation speed of chitin.
The preparation method of the controllable labyrinth chitosan oligosaccharide of the degree of polymerization and deacetylation is added in faintly acid solvent
Hydrochloric acid or sulfuric acid quality be solvent quality 0.2~12%.
The preparation method of the controllable labyrinth chitosan oligosaccharide of the degree of polymerization and deacetylation, the alcohol precipitation that alcohol deposition method uses
Reagent is one or more of methanol, ethyl alcohol, propyl alcohol, isopropanol, butanol, isoamyl alcohol, acetone, butanone.
The preparation method of the controllable labyrinth chitosan oligosaccharide of the degree of polymerization and deacetylation, further regulates and controls chitosan oligosaccharide
Non-specificity commercial enzyme used in the degree of polymerization and deacetylation, using cellulase, protease, amylase or lipase.
Design philosophy of the invention is:
By the research of over half a century, it has been found that a variety of solution systems that can dissolve chitin, including high concentration
Strong acid, trichloroacetic acid, dichloroacetic acid, formic acid, methane sulfonic acid, the chlorhydrin etc. such as hydrochloric acid, nitric acid, sulfuric acid, phosphoric acid it is highly polar molten
Agent, amide/lithium chloride system, fluorated solvent etc..The invention firstly uses dense strong acid, intensive polar solvent, fluorated solvent or amide/
The dissolution such as lithium chloride solvent is sufficiently swollen chitin, and the chitin after dissolution or sufficiently swelling is degraded under acid catalysed conditions
Small molecule chitin or chitin oligo saccharide.This degradation process is able to achieve the controlled degradation of chitin, and obtaining molecular weight can be in number
Thousand to be continuously adjusted in tens of thousands of ranges, small molecule chitin or chitin oligo saccharide of the breadth coefficient pid value less than 1.7.
People have found chitinase, chitosan enzyme, chitin deacetylase and are contained in Enzymatic Hydrolysis of Chitosan technical research
The commodity enzyme of above-mentioned enzymolysis activity has certain degradation to chitin or its different degrees of deacetylated product chitosan
Activity, and activity and the structure of product and the deacetylation of substrate are directly related.The present invention drops acid using these hydrolases
It solves the small molecule chitin generated or chitin oligo saccharide carries out further structure regulating, to realize final product labyrinth shell
The Effective Regulation of the oligosaccharides degree of polymerization (less than 20) and deacetylation (20%~80%).
The invention has the advantages and beneficial effects that:
1, the present invention uses makes chitin dissolve in reaction dissolvent or sufficiently be swollen first, then the even phase drop under acid catalysis
The method of solution carries out reaction under conditions of milder, and reaction process is easily controllable, and can effectively control small molecule crust
The degree of polymerization distribution of element or chitin oligo saccharide product.
2, the present invention proposes using chitinase, chitosan enzyme, chitin deacetylase or contains above-mentioned enzymolysis activity
The commercial enzyme structure of small molecule chitin or chitin oligo saccharide that acid degradation is generated carry out deeper into regulation.
3, the preparation method of heretofore described labyrinth chitosan oligosaccharide can be realized to the chitosan oligosaccharide degree of polymerization and de- second
The Effective Regulation of the sugar chain molecules structure such as acyl degree, production process high income, by-product is few, at low cost, is easy to mass production.
Detailed description of the invention
Fig. 1 is acid degradation product chitin oligo saccharide MALDI-TOF mass spectrogram.In figure, abscissa represents mass-to-charge ratio (m/z), indulges
Coordinate represents quasi-molecular ions signal strength.
Fig. 2 is acid degradation product chitin oligo saccharide1H NMR spectra.In figure, abscissa represents chemical shift, and ordinate represents
Spectral peak signal strength.
Fig. 3 is chitin oligo saccharide group synthase (S+C) catabolite MALDI-TOF mass spectrogram.In figure, abscissa represents mass-to-charge ratio
(m/z), ordinate represents quasi-molecular ions signal strength.
Fig. 4 is chitin oligo saccharide group synthase (S+C) catabolite1H NMR spectra.In figure, abscissa represents chemical shift, indulges
Coordinate represents spectral peak signal strength.
Fig. 5 is chitin oligo saccharide commercial fibre element enzyme catabolite MALDI-TOF mass spectrogram.In figure, abscissa represents matter lotus
Than (m/z), ordinate represents quasi-molecular ions signal strength.
Fig. 6 is chitin oligo saccharide commercial fibre element enzyme catabolite1H NMR spectra.In figure, abscissa represents chemical shift,
Ordinate represents spectral peak signal strength.
Fig. 7 is the deacetylated product MALDI-TOF mass spectrogram of chitin oligo saccharide chitin deacetylase.In figure, abscissa is represented
Mass-to-charge ratio (m/z), ordinate represent quasi-molecular ions signal strength.
Fig. 8 is the deacetylated product of chitin oligo saccharide chitin deacetylase1H NMR spectra.In figure, abscissa represents chemical potential
It moves, ordinate represents spectral peak signal strength.
Fig. 9 is different small molecule chitins, chitin oligo saccharide and chitosan oligosaccharide molecular-exclusion chromatography figure.In figure, abscissa is represented
Retention time (minute), ordinate represent detection electric signal.
Specific embodiment
In the specific implementation process, the preparation method of labyrinth chitosan oligosaccharide of the present invention, includes the following steps:
(1) sheet chitin is crushed to appropriate granularity with pulverizer, generally about (80~200 mesh).
(2) crust crude granule is scattered in 5 times to 50 times solvents, under strong stirring, chitin dissolve in a solvent or
Sufficiently swelling.
The solvent for dissolving chitin is hexafluoroisopropanol, Hexafluoro acetone, dichlroroethanol, the dimethyl formyl containing lithium chloride
Amine, the dimethyl acetamide containing lithium chloride, N- methyl morpholine oxide, concentrated hydrochloric acid, the concentrated sulfuric acid, concentrated phosphoric acid, methane sulfonic acid, first
The mixture of one or more of acid, dichloroacetic acid, trichloroacetic acid, solvent usage are 5~50 times of chitin quality.
When using faintly acid solvent: hexafluoroisopropanol, Hexafluoro acetone, dichlroroethanol, the dimethyl formyl containing lithium chloride
Amine, the dimethyl acetamide containing lithium chloride, the one or two of N- methyl morpholine oxide, formic acid, dichloroacetic acid, trichloroacetic acid
When above, by the way that hydrochloric acid or sulfuric acid is added, to improve the degradation speed of chitin.
The hydrochloric acid or sulfuric acid quality being added in faintly acid solvent are the 0.2~12% of solvent quality.
(3) reactant keeps the temperature 1 to 10 hour at 0 DEG C~100 DEG C, and chitin starts to degrade, and molecular weight gradually reduces, and passes through
Reaction temperature and reaction time are controlled, the small molecule chitin or chitin oligo saccharide that there is different polymerization degree to be distributed are obtained.
(4) reaction liquid solvent is removed using the methods of vacuum decompression evaporation solvent, alcohol precipitation.
(5) chitinase, chitosan enzyme, chitin deacetylase or the non-specificity with above-mentioned enzymolysis activity are used
Commodity enzyme further regulates and controls the degree of polymerization and deacetylation of small molecule chitin or chitin oligo saccharide, obtains with different molecular knot
The complicated chitosan oligosaccharide of structure.
In order to keep technical solution of the present invention and advantage clearer, retouched in detail below in conjunction with specific embodiment
It states.
Embodiment 1
At 10 DEG C, 10g chitin is slowly added into 200mL concentrated hydrochloric acid (36wt%).After stirring 6 hours, chitin
It is completely dissolved.Reaction temperature is improved to 30 DEG C, is stirred 1 hour, and is slowly added to the neutralization of NaOH solid powder in backward reaction solution
Reaction solution.Reaction solution is added 1000mL ethyl alcohol and carries out alcohol precipitation, and the solid precipitating of precipitation is filtered, ethyl alcohol cleaning, and air blast is dry
It is dry to obtain 6 grams of small molecule chitins and chitin oligo saccharide.
Embodiment 2
At 30 DEG C, 10g chitin is slowly added into 500mL hexafluoroisopropanol.Stirring 6 hours, chitin is completely molten
25mL concentrated hydrochloric acid is added after solution.Reaction temperature is improved to 50 DEG C, after stirring 1.2 hours, is rotated in 50 DEG C of vacuum, makes the overwhelming majority
Reaction dissolvent evaporation recycling.Ethyl alcohol cleaning is added in reaction product, and filtering, it is few that forced air drying obtains 7 grams of small molecule chitin chitins
Sugared mixture.
Embodiment 3
At 20 DEG C, 10g chitin is slowly added into 300mL formic acid.Stirring 6 hours, chitin is added after being completely dissolved
21mL concentrated hydrochloric acid.Reaction temperature is improved to 64 DEG C, after stirring 1.2 hours, is rotated in 50 DEG C of vacuum, makes most reaction dissolvents
Evaporation recycling.Ethyl alcohol cleaning, filtering is added in reaction product, and forced air drying obtains 7 grams of small molecule chitins and chitin oligo saccharide.
Embodiment 4
In the present embodiment, the degree of polymerization and the controllable labyrinth chitosan oligosaccharide of deacetylation the preparation method is as follows:
At 20 DEG C, 10g chitin is slowly added into 300mL dichloroacetic acid.Stirring 6 hours, after chitin is completely dissolved
5mL concentrated hydrochloric acid is added.Reaction temperature is improved to 70 DEG C, after stirring 1.2 hours, is rotated in 60 DEG C of vacuum, makes most reactions
Solvent evaporation recycling.Ethyl alcohol cleaning, filtering is added in reaction product, and forced air drying obtains 8 grams of small molecule chitins and chitin oligo saccharide.
It is first analyzed using composition of the MALDI-TOF MS to product, the results show that product is mainly the acetylation of the degree of polymerization 2~7
Chitin oligo saccharide (Fig. 1)；In addition, passing through1H NMR is measured the deacetylation of product, the results show that product chitosan oligosaccharide is de-
Acetyl degree is 4%, and be consistent (Fig. 2) with mass spectral results.
Embodiment 5
In the present embodiment, the degree of polymerization and the controllable labyrinth chitosan oligosaccharide of deacetylation the preparation method is as follows:
Neutral small molecule chitin/chitin oligo saccharide 1g from embodiment 4 is weighed, 1% aqueous solution is configured to, table is added
The chitinase (S) and chitosan enzyme (C) each 1mg reached is combined degradation, and reaction temperature is 40 DEG C, and the reaction time is for 24 hours.Make
It is analyzed with composition of the MALDI-TOF MS to product, the results show that including different deacetylations, the degree of polymerization 2~7 in product
Chitosan oligosaccharide (Fig. 3)；In addition, passing through1H NMR is measured the overall deacetylation of product chitosan oligosaccharide, the results show that product
The deacetylation of labyrinth chitosan oligosaccharide is 26% (Fig. 4).
Embodiment 6
In the present embodiment, the degree of polymerization and the controllable labyrinth chitosan oligosaccharide of deacetylation the preparation method is as follows:
Neutral small molecule chitin/chitin oligo saccharide 1g from embodiment 4 is weighed, 1% aqueous solution is configured to, quotient is added
Product cellulase 0.1g degrades, and reaction temperature is 40 DEG C, and the reaction time is for 24 hours.Using MALDI-TOF MS to product
Composition is analyzed, the results show that including different deacetylations, the chitosan oligosaccharide (Fig. 5) of the degree of polymerization 2~8 in product；In addition, logical
It crosses 1H NMR to be measured the overall deacetylation of product chitosan oligosaccharide, the results show that the de- second of product labyrinth chitosan oligosaccharide
Acyl degree is 26% (Fig. 6).
Embodiment 7
In the present embodiment, the degree of polymerization and the controllable labyrinth chitosan oligosaccharide of deacetylation the preparation method is as follows:
Neutral small molecule chitin/chitin oligo saccharide 1g from embodiment 4 is weighed, 1% aqueous solution is configured to, table is added
The chitin deacetylase freeze-dried powder 1mg reached degrades, and reaction temperature is 40 DEG C, and the reaction time is for 24 hours.Use MALDI-
TOF MS analyzes the composition of product, the results show that product includes different deacetylations, the chitosan oligosaccharide of the degree of polymerization 2~10
(Fig. 7)；In addition, being measured by deacetylation of the 1H NMR to product chitosan oligosaccharide, the results show that the de- second of product chitosan oligosaccharide
Acyl degree is 50% (Fig. 8).The small molecule chitin that using Size Exclusion High Performance liquid-phase chromatography method prepared by above-mentioned 7 examples,
Chitin oligo saccharide and chitosan oligosaccharide product carry out molecular weight distribution analysis, concrete outcome as shown in figure 9, product molecular weight 1500~
It is continuously adjusted in 36000.
Embodiment the result shows that, preparation method of the present invention can be realized to the degree of polymerization of complicated chitosan oligosaccharide and deacetylation etc.
Sugar chain molecule structure is regulated and controled, and product yield is high, and by-product is few, at low cost, is easy to mass production.
Claims (8)
1. the preparation method of a kind of degree of polymerization and the controllable labyrinth chitosan oligosaccharide of deacetylation, which is characterized in that make first first
Shell element dissolves in a solvent or sufficiently swelling, and even phase degrades to obtain small molecule chitin or chitin oligo saccharide under acid catalysis, and small point
Sub- chitin or chitin oligo saccharide chitinase, chitosan enzyme, chitin deacetylase contain the non-of above-mentioned enzymolysis activity
Specific commercial enzyme further regulates and controls the degree of polymerization and deacetylation, spray-dried to obtain the degree of polymerization 20 hereinafter, deacetylated
Degree regulatable labyrinth chitosan oligosaccharide between 20~80%.
2. the preparation method of the degree of polymerization according to claim 1 and the controllable labyrinth chitosan oligosaccharide of deacetylation, special
Sign is, the specific steps are as follows:
1) it is scattered in after chitin is crushed with pulverizer in the reaction dissolvent of 5 times to 50 times quality, under strong stirring, chitin
It is sufficiently swollen or dissolves in reaction dissolvent；
2) reactant keeps the temperature 1 to 10 hour at 0 DEG C~100 DEG C, chitin even phase degradation under acid catalysis, when the degree of polymerization is with reaction
Between gradually reduce, until obtain small molecule chitin or chitin oligo saccharide solution；
3) reaction dissolvent is removed using the evaporation of vacuum decompression solvent, alcohol precipitation and vacuum drying, spray drying or forced air drying, and done
It is dry to obtain small molecule chitin or chitin oligo saccharide；
4) small molecule chitin or chitin oligo saccharide chitinase, chitosan enzyme, chitin deacetylase or contain above-mentioned enzyme
Solve the degree of polymerization and deacetylation that active non-specific commercial enzyme further regulates and controls oligosaccharides；Spray-dried processing is gathered
It is right 20 hereinafter, deacetylation regulatable labyrinth chitosan oligosaccharide between 20~80%.
3. the preparation method of the degree of polymerization according to claim 1 or 2 and the controllable labyrinth chitosan oligosaccharide of deacetylation,
It is characterized in that, chitin is crushed to 80~200 mesh after rinsing drying, then using pulverizer.
4. the preparation method of the degree of polymerization according to claim 1 or 2 and the controllable labyrinth chitosan oligosaccharide of deacetylation,
It is characterized in that, the solvent for dissolving chitin is hexafluoroisopropanol, Hexafluoro acetone, dichlroroethanol, the dimethyl methyl containing lithium chloride
Amide, the dimethyl acetamide containing lithium chloride, N- methyl morpholine oxide, concentrated hydrochloric acid, the concentrated sulfuric acid, concentrated phosphoric acid, methane sulfonic acid, first
The mixture of one or more of acid, dichloroacetic acid, trichloroacetic acid, solvent usage are 5~50 times of chitin quality.
5. the preparation method of the degree of polymerization according to claim 4 and the controllable labyrinth chitosan oligosaccharide of deacetylation, special
Sign is, when using faintly acid solvent: hexafluoroisopropanol, Hexafluoro acetone, dichlroroethanol, the dimethyl formyl containing lithium chloride
Amine, the dimethyl acetamide containing lithium chloride, the one or two of N- methyl morpholine oxide, formic acid, dichloroacetic acid, trichloroacetic acid
When above, by the way that hydrochloric acid or sulfuric acid is added, to improve the degradation speed of chitin.
6. the preparation method of the degree of polymerization according to claim 5 and the controllable labyrinth chitosan oligosaccharide of deacetylation, special
Sign is that the hydrochloric acid or sulfuric acid quality being added in faintly acid solvent are the 0.2~12% of solvent quality.
7. the preparation method of the degree of polymerization according to claim 2 and the controllable labyrinth chitosan oligosaccharide of deacetylation, special
Sign is that the alcohol precipitation reagent that alcohol deposition method uses is in methanol, ethyl alcohol, propyl alcohol, isopropanol, butanol, isoamyl alcohol, acetone, butanone
It is one or more kinds of.
8. the preparation method of the degree of polymerization according to claim 2 and the controllable labyrinth chitosan oligosaccharide of deacetylation, special
Sign is, further regulates and controls non-specificity commercial enzyme used in the chitosan oligosaccharide degree of polymerization and deacetylation, using cellulase, egg
White enzyme, amylase or lipase.
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| CN114058659A (en) * | 2021-11-17 | 2022-02-18 | 河北农业大学 | Preparation method of chitin oligosaccharide |
| CN114965543A (en) * | 2022-06-13 | 2022-08-30 | 青岛科技大学 | Method for determining attribution of chitosan oligosaccharide NMR spectrum and related index of deacetylation degree |
| CN114965543B (en) * | 2022-06-13 | 2023-10-20 | 青岛科技大学 | Determination method of chitosan oligosaccharide NMR spectrum attribution and deacetylation degree related index |
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