CN102492664A - Method for preparing chitosan oligosaccharide by applying complex enzyme - Google Patents
Method for preparing chitosan oligosaccharide by applying complex enzyme Download PDFInfo
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- CN102492664A CN102492664A CN2011104052849A CN201110405284A CN102492664A CN 102492664 A CN102492664 A CN 102492664A CN 2011104052849 A CN2011104052849 A CN 2011104052849A CN 201110405284 A CN201110405284 A CN 201110405284A CN 102492664 A CN102492664 A CN 102492664A
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- prozyme
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- oligochitosan
- chitosan
- enzyme
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- 108090000790 Enzymes Proteins 0.000 title claims abstract description 37
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 27
- RQFQJYYMBWVMQG-IXDPLRRUSA-N chitotriose Chemical compound O[C@@H]1[C@@H](N)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](N)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)N)[C@@H](CO)O1 RQFQJYYMBWVMQG-IXDPLRRUSA-N 0.000 title abstract 4
- 229920001661 Chitosan Polymers 0.000 claims abstract description 99
- 229940088598 enzyme Drugs 0.000 claims abstract description 36
- 108010064696 N,O-diacetylmuramidase Proteins 0.000 claims description 12
- 229920001353 Dextrin Polymers 0.000 claims description 11
- 239000004375 Dextrin Substances 0.000 claims description 11
- 235000019425 dextrin Nutrition 0.000 claims description 11
- 239000012043 crude product Substances 0.000 claims description 10
- 239000002808 molecular sieve Substances 0.000 claims description 10
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 claims description 10
- 108010059892 Cellulase Proteins 0.000 claims description 9
- 229940106157 cellulase Drugs 0.000 claims description 9
- 235000019154 vitamin C Nutrition 0.000 claims description 9
- 239000011718 vitamin C Substances 0.000 claims description 9
- 239000000126 substance Substances 0.000 claims description 8
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 7
- 235000017281 sodium acetate Nutrition 0.000 claims description 7
- 239000001632 sodium acetate Substances 0.000 claims description 7
- 239000002131 composite material Substances 0.000 claims description 5
- 239000000706 filtrate Substances 0.000 claims description 5
- 108010084185 Cellulases Proteins 0.000 claims description 4
- 102000005575 Cellulases Human genes 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims 2
- KTVIXTQDYHMGHF-UHFFFAOYSA-L cobalt(2+) sulfate Chemical compound [Co+2].[O-]S([O-])(=O)=O KTVIXTQDYHMGHF-UHFFFAOYSA-L 0.000 claims 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims 1
- 235000019341 magnesium sulphate Nutrition 0.000 claims 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims 1
- 229910000368 zinc sulfate Inorganic materials 0.000 claims 1
- 239000011686 zinc sulphate Substances 0.000 claims 1
- 235000009529 zinc sulphate Nutrition 0.000 claims 1
- 239000000243 solution Substances 0.000 abstract description 17
- 238000009826 distribution Methods 0.000 abstract description 4
- 239000011535 reaction buffer Substances 0.000 abstract description 2
- 239000004382 Amylase Substances 0.000 abstract 1
- 102000013142 Amylases Human genes 0.000 abstract 1
- 108010065511 Amylases Proteins 0.000 abstract 1
- 239000004367 Lipase Substances 0.000 abstract 1
- 102000004882 Lipase Human genes 0.000 abstract 1
- 108090001060 Lipase Proteins 0.000 abstract 1
- 102000016943 Muramidase Human genes 0.000 abstract 1
- 108010014251 Muramidase Proteins 0.000 abstract 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 abstract 1
- 108090000526 Papain Proteins 0.000 abstract 1
- 239000004365 Protease Substances 0.000 abstract 1
- 235000019418 amylase Nutrition 0.000 abstract 1
- 229920002678 cellulose Polymers 0.000 abstract 1
- 239000001913 cellulose Substances 0.000 abstract 1
- 235000010980 cellulose Nutrition 0.000 abstract 1
- 235000019421 lipase Nutrition 0.000 abstract 1
- 229960000274 lysozyme Drugs 0.000 abstract 1
- 239000004325 lysozyme Substances 0.000 abstract 1
- 235000010335 lysozyme Nutrition 0.000 abstract 1
- 229940055729 papain Drugs 0.000 abstract 1
- 235000019834 papain Nutrition 0.000 abstract 1
- 238000002360 preparation method Methods 0.000 description 13
- 238000006116 polymerization reaction Methods 0.000 description 12
- 230000015556 catabolic process Effects 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 7
- 238000006731 degradation reaction Methods 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 150000003839 salts Chemical class 0.000 description 6
- 239000002994 raw material Substances 0.000 description 5
- 230000006196 deacetylation Effects 0.000 description 4
- 238000003381 deacetylation reaction Methods 0.000 description 4
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000002950 deficient Effects 0.000 description 2
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 108010059820 Polygalacturonase Proteins 0.000 description 1
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 230000002478 diastatic effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 229960002442 glucosamine Drugs 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229920000550 glycopolymer Polymers 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- GHMLBKRAJCXXBS-UHFFFAOYSA-N resorcinol Chemical compound OC1=CC=CC(O)=C1 GHMLBKRAJCXXBS-UHFFFAOYSA-N 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 239000010454 slate Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- -1 zytase Proteins 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a method for preparing a chitosan oligosaccharide by applying a complex enzyme. The complex enzyme comprises cellulose, lysozyme, amylase, lipase, glucolase and papain. The invention further provides a method for preparing a chitosan oligosaccharide by applying the complex enzyme. The method comprises the following step of: adding the complex enzyme into a reaction buffer solution of chitosan, wherein the mass ratio of the complex enzyme to the chitosan is (1-20):100, and is preferably (5-15):100. The complex enzyme and the chitosan oligosaccharide prepared from the complex enzyme have uniform molecular weight distribution and high yields.
Description
Technical field
The invention belongs to oligose preparation method field, relate in particular to the method that a kind of applying composite enzyme prepares oligochitosan.
Background technology
Oligochitosan is very valuable natural polysaccharide, and it has broad application prospects in medicine and agricultural, and still for the preparation of oligochitosan, especially the applying biological legal system is equipped with oligochitosan, exists process complicated, and degradation efficiency is low, characteristics such as product slate variation.
The scheme that for example present applying composite enzyme prepares oligochitosan mainly is the prozyme that utilizes glycase, proteolytic enzyme and Remazol Brilliant Violet carboxymethyl chitin hydrolase etc. to form according to certain ratio; Like disclosed compound enzymic preparation among the patented claim CN101200711A; It is the enzyme that obtains through fermentation of bacillus subtilis; The kind of enzyme and content receive in the microbe body had the quantity limitation of enzyme, so the efficient of degraded is not high.
Be exactly to use a kind of non-specificity enzyme to prepare oligochitosan in addition; For example disclosed application papoid prepares oligochitosan among the patent CN101475969A; But papoid efficient in the chitosan of degraded high-polymerization degree is high; Degradation efficiency for the chitosan of low polymerization degree is lower, so the selectivity of its degraded is single.
Disclosed among the patent CN101818181A in addition, application polygalacturonase, zytase, cellulase, diastatic prozyme prepare oligochitosan, but time of its preparation oligochitosan is long, and the oligochitosan color that obtains is darker.
At present also useful physics method, chemical method wait degrade chitosan to prepare oligochitosan, otherwise since degrade not thorough, make to have too much high-polymerization degree oligochitosan in the product; For example the polymerization degree occupies the majority greater than 50 chitosan oligomer; Or degrade excessively, make to have too much glucosamine sugar monomer in the product, or the polymerization degree is that the oligochitosan of 2-10 is too much; In order to obtain polymerization degree height bonded oligochitosan, there is certain defective in present preparation technology.
Summary of the invention
For solving the problems of the technologies described above; The invention provides a kind of degrade chitosan prepares the prozyme of oligochitosan and uses the method that this prozyme prepares oligochitosan; This prozyme can make that the control of the oligochitosan polymerization degree within the specific limits in the degraded product; Just the polymerization degree is controlled in 2-30 (molecular weight the is 300-5000D) scope, and the oligochitosan of different polymerization degree evenly exists in product, and this oligochitosan makes in the agricultural application more efficient.
Particularly, the invention provides a kind of prozyme for preparing oligochitosan, this mixture contains the enzyme of following mass fraction:
Cellulase 15-25 part,
N,O-Diacetylmuramidase 5-15 part
Glycase 20-40 part
Lypase 80-100 part
Dextrin 20-30 part
Papoid 10-20 part.
Preferably, above-mentioned prozyme contains the enzyme of following mass fraction:
20 parts of cellulases,
10 parts of N,O-Diacetylmuramidases
30 parts of glycase
90 parts in lypase
25 parts of dextrins
15 parts of papoids.
In the above-mentioned prozyme of the present invention, can comprise the material that other promotion enzyme is lived, vitamin substances for example, like vitamins C, or metal-salt, like CoSO
4, MnSO
4, FeSO
4, MgSO
4, ZnSO
4, K
2SO
4Deng, add the decomposition rate that these materials can promote enzyme.
Preferably, also contain vitamins C in the above-mentioned prozyme of the present invention, the mass ratio of vitamins C and prozyme is 0.5-4: 1.Perhaps preferred, be 1.5-3: 1, preferred, also contain CoSO in the above-mentioned prozyme of the present invention
4, MnSO
4, FeSO
4, MgSO
4, ZnSO
4, K
2SO
4In one or more, no matter comprise several kinds of metal-salts, the molar weight of various metal-salts is identical, and is preferred, comprises wherein 6 kinds of salt simultaneously, the molar weight of 6 kinds of salt is identical, the total mass of 6 kinds of salt is 1-10 with the ratio of the total mass of enzyme: 100, preferred 1-5: 100.
The present invention also provides the method that above-mentioned prozyme prepares oligochitosan of using; This method is included in the reaction buffer of chitosan and adds prozyme; Make that the mass ratio of prozyme and chitosan is 1-20: 100, preferred, the mass ratio of prozyme and chitosan is 5-15: 100.
Concrete, the invention provides a kind of method that above-mentioned prozyme prepares oligochitosan of using, this method comprises the steps:
1) chitosan is dissolved with acetum, use sodium acetate soln to transfer pH to be 5-6 then, be mixed with chitosan concentration, get chitosan solution at 50-70g/L;
2) in the chitosan solution that step 1) makes, add prozyme, the component and the mass fraction thereof of prozyme are following:
Cellulase 15-25 part,
N,O-Diacetylmuramidase 5-15 part
Glycase 20-40 part
Lypase 80-100 part
Dextrin 20-30 part
Papoid 10-20 part.
The mass ratio of prozyme and chitosan is: 1-20: 100, and temperature 50-60 ℃ of shaking table hunting speed degraded 2-3 hour under 50-100 rev/min the condition, prepare the oligochitosan crude product solution;
3) with the molecular sieve of oligochitosan crude product solution through 5000D, filtrate through the molecular sieve of 300D, with the trapped substance lyophilize, prepares oligochitosan again.
In the aforesaid method, because prozyme of the present invention can be applicable to the degraded of high deacetylized chitosan, also applicable to the degraded of low deacetylation chitosan, so the deacetylation of chitosan does not generally have special limitation.General, the deacetylation of the raw materials of chitosan that above-mentioned preparation method of the present invention is used is 60-99%.
In the aforesaid method, the concentration of used acetum is unrestricted, and general concentration is 0.6mol/L, and the concentration of sodium acetate soln is also unrestricted, and general concentration is 0.6mol/L.
Preferably, in the aforesaid method, prozyme is made up of the enzyme of following mass fraction:
20 parts of cellulases,
10 parts of N,O-Diacetylmuramidases
30 parts of glycase
90 parts in lypase
25 parts of dextrins
15 parts of papoids.
Preferably, in the aforesaid method, the mass ratio of prozyme and chitosan is 5-15: 100.
Preferably, in the aforesaid method, when adding prozyme, can add a certain proportion of vitamins C, the mass ratio of vitamins C and prozyme is 0.5-4: 1.Perhaps preferred, be 1.5-3: 1, in the aforesaid method, when adding prozyme, can add CoSO
4, MnSO
4, FeSO
4, MgSO
4, ZnSO
4, K
2SO
4In one or more, no matter add several kinds of metal-salts, the molar weight of various metal-salts is identical, and is preferred, comprises wherein 6 kinds of salt simultaneously, the molar weight of 6 kinds of salt is identical, the total mass of 6 kinds of salt is 1-10 with the ratio of the total mass of enzyme: 100, preferred 1-5: 100.
Contain multiple non-narrow spectrum degrading enzyme in the prozyme of the present invention; Wherein N,O-Diacetylmuramidase can be degraded owing to take off the incomplete chitosan of acetyl; Just cut the n acetylglucosamine n glycopolymers, and cellulase can arrive lower molecular weight with degradation of chitosan owing to be restriction endonuclease.Papoid can be with the degradation of chitosan of high-polymerization degree, and degradation efficiency is high.Above-mentioned several kinds of enzymes are mixed; Make the efficient of degraded improve; Can both have effective degraded to the chitosan of various polymerization thing degree, the chitosan that takes off acetyl and do not take off acetyl etc.; Avoided the defective of single kind of non-specificity degradation of chitosan, and the set through different non-specificity enzymes, made that the GS polymkeric substance of different polymerization degree evenly exists in the product that degraded obtains; And range of molecular weight distributions is that the oligochitosan ratio of 1000-5000 is high, and the productive rate of preparation target oligochitosan is high.
Use the active unaffected of oligochitosan that the enzymic degradation chitosan obtains, generally keep natural oligochitosan structure, so in medical and agricultural application, have good effect.And the oligochitosan even molecular weight distribution for preparing of prozyme of the present invention and applying composite enzyme thereof, productive rate is high.
Embodiment
In order to understand the present invention, further specify the present invention with embodiment below, but do not limit the present invention.
Used raw material all is to buy the raw material that obtains from market among the embodiment, and the deacetylation of chitosan is 85%, and other raw material is analytically pure raw material.
Embodiment 1: the preparation of prozyme I
With cellulase 20g, N,O-Diacetylmuramidase 10g, glycase 30g, lypase 90g, dextrin 25g, papoid 15g mixes, and prepares prozyme I.
Embodiment 2: the preparation of compound enzyme II
With cellulase 25g, N,O-Diacetylmuramidase 5g, glycase 20g, lypase 90g, dextrin 30g, papoid 10g mixes, and adds the vitamins C of 317g, prepares compound enzyme II.
Embodiment 3: the preparation of compound enzyme II I
With cellulase 15g, N,O-Diacetylmuramidase 15g, glycase 40g, lypase 80g, dextrin 20g, papoid 20g mixes, and adds the CoSO of 0.155g
4, 0.151g MnSO
4, 0.152g FeSO
4, 0.12g MgSO
4, 0.161g ZnSO
4, 0.174g K
2SO
4, prepare compound enzyme II I.
The preparation of embodiment 4 oligochitosans
1) with of the acetum dissolving of 100g chitosan, transfers to pH=5.8 with the 0.6mol/L sodium acetate soln then, be mixed with the chitosan solution of 2L with 0.6mol/L;
2) the prozyme I that the embodiment 1 of adding 15g prepares in chitosan solution.Be degraded 3 hours under 70 rev/mins the condition in 55 ℃ of shaking table hunting speeds of temperature, prepare the oligochitosan crude product solution;
3) with the molecular sieve of oligochitosan crude product solution through 5000D, filtrate through the molecular sieve of 300D, with the trapped substance lyophilize, prepares the 87.4g oligochitosan again.
The preparation of embodiment 5 oligochitosans
1) with of the acetum dissolving of 100g chitosan, transfers to pH=5.9 with the 0.6mol/L sodium acetate soln then, be mixed with the chitosan solution of 2L with 0.6mol/L;
2) compound enzyme II that the embodiment 2 of adding 12g prepares in chitosan solution.Be degraded 2.5 hours under 80 rev/mins the condition in 57 ℃ of shaking table hunting speeds of temperature, prepare the oligochitosan crude product solution;
3) with the molecular sieve of oligochitosan crude product solution through 5000D, filtrate through the molecular sieve of 300D, with the trapped substance lyophilize, prepares the 89.6g oligochitosan again.
The preparation of embodiment 6 oligochitosans
1) with of the acetum dissolving of 100g chitosan, transfers to pH=5.7 with the 0.6mol/L sodium acetate soln then, be mixed with the chitosan solution of 2L with 0.6mol/L;
2) the compound enzyme II I that the embodiment 3 of adding 10g prepares in chitosan solution.Be degraded 2 hours under 90 rev/mins the condition in 58 ℃ of shaking table hunting speeds of temperature, prepare the oligochitosan crude product solution;
3) with the molecular sieve of oligochitosan crude product solution through 5000D, filtrate through the molecular sieve of 300D, with the trapped substance lyophilize, prepares the 90.1g oligochitosan again.
The MWD of the oligochitosan that embodiment 4,5 and 6 prepares is 300-5000D; And productive rate is greater than 85%; This oligochitosan separates through molecular-weight gradation, records oligochitosan in the different molecular weight section and distributes comparatively evenly, the oligochitosan that the high oligochitosan of molecular weight occupies the majority or molecular weight is low do not occur and occupies the majority; That is to say the oligochitosan even molecular weight distribution that process prozyme of the present invention prepares.And use the time weak point that method of the present invention prepares oligochitosan, be generally the degraded that can realize oligochitosan in 2-3 hour.
It can also be seen that through the foregoing description, added the active promoted material of prozyme, vitamins C and metal-salt etc. have not only reduced the consumption of prozyme, and the shortening of DeR time, and the productive rate of the product of reaction improves on the contrary.
Claims (8)
1. prozyme for preparing oligochitosan, this mixture comprises the enzyme of following mass fraction:
Cellulase 15-25 part,
N,O-Diacetylmuramidase 5-15 part
Glycase 20-40 part
Lypase 80-100 part
Dextrin 20-30 part
Papoid 10-20 part.
2. prozyme according to claim 1, said prozyme comprises the enzyme of following mass fraction:
20 parts of cellulases,
10 parts of N,O-Diacetylmuramidases
30 parts of glycase
90 parts in lypase
25 parts of dextrins
15 parts of papoids.
3. prozyme according to claim 1 also contains vitamins C in this prozyme, the mass ratio of vitamins C and prozyme is 0.5-4: 1.
4. prozyme according to claim 1 also contains metal-salt CoSO4, MnSO4, FeSO4, MgSO4, ZnSO4 and K in this prozyme
2SO4, the molar weight of various metal-salts is identical, and the total mass of metal-salt is 1-10 with the ratio of the total mass of enzyme: 100.
5. an applying composite enzyme prepares the method for oligochitosan, and this method comprises the steps:
1) chitosan is dissolved with acetum, use sodium acetate soln to transfer pH to be 5-6 then, be mixed with chitosan concentration, get chitosan solution at 50-70g/L;
2) in the chitosan solution that step 1) makes, add prozyme, the component and the mass fraction thereof of prozyme are following:
Cellulase 15-25 part,
N,O-Diacetylmuramidase 5-15 part
Glycase 20-40 part
Lypase 80-100 part
Dextrin 20-30 part
Papoid 10-20 part,
The mass ratio of prozyme and chitosan is: 1-20: 100, and temperature 50-60 ℃ of shaking table hunting speed degraded 2-3 hour under 50-100 rev/min the condition, prepare the oligochitosan crude product solution;
3) with the molecular sieve of oligochitosan crude product solution through 5000D, filtrate through the molecular sieve of 300D, with the trapped substance lyophilize, prepares oligochitosan again.
6. method according to claim 5, wherein the concentration of acetum is 0.6mol/L, the concentration of sodium acetate soln is 0.6mol/L.
7. method according to claim 5, wherein prozyme comprises the enzyme of following mass fraction:
20 parts of cellulases,
10 parts of N,O-Diacetylmuramidases
30 parts of glycase
90 parts in lypase
25 parts of dextrins
15 parts of papoids.
8. method according to claim 5, wherein the mass ratio of prozyme and chitosan is 5-15: 100.
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|---|---|---|---|
| CN2011104052849A CN102492664B (en) | 2011-12-08 | 2011-12-08 | Method for preparing chitosan oligosaccharide by applying complex enzyme |
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|---|---|---|---|
| CN2011104052849A CN102492664B (en) | 2011-12-08 | 2011-12-08 | Method for preparing chitosan oligosaccharide by applying complex enzyme |
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| CN102492664B CN102492664B (en) | 2013-01-09 |
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| CN103146784A (en) * | 2013-01-16 | 2013-06-12 | 江南大学 | Method for preparing chitosan oligosaccharide through degradation of chitosan by thermobifida fusca internally tangent beta-1, 4-glucanase |
| CN103689112A (en) * | 2013-12-06 | 2014-04-02 | 柳州联海科技有限公司 | Preparation method of chitooligosaccharide-containing beancurd |
| JP2015059071A (en) * | 2013-09-19 | 2015-03-30 | 秀夫 草桶 | Fertilizer containing chitin and/or chitosan or chitin and/or chitosan inclusion and culture of microorganisms having chitin and/or chitosan resolving ability, and production method thereof |
| CN105803019A (en) * | 2015-12-31 | 2016-07-27 | 珠海市金隆生物科技有限公司 | Method for producing chitosan oligosaccharide from home-made enzyme solution |
| CN106755208A (en) * | 2016-12-30 | 2017-05-31 | 山东智顺进出口有限公司 | The method that low energy consumption digests shitosan preparation solution body shell oligosaccharides |
| CN107602727A (en) * | 2017-11-06 | 2018-01-19 | 贾晗 | A kind of sugared preparation method to eight sugared chromic compounds of shell six |
| CN109438103A (en) * | 2019-01-10 | 2019-03-08 | 江苏心实肥业集团有限公司 | A kind of chitosan oligosaccharide crop nutrition spray fertilizer and preparation method thereof |
| CN109984130A (en) * | 2017-12-31 | 2019-07-09 | 海南正业中农高科股份有限公司 | A kind of chitosan oligosaccharide composition and application thereof |
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| CN103146784A (en) * | 2013-01-16 | 2013-06-12 | 江南大学 | Method for preparing chitosan oligosaccharide through degradation of chitosan by thermobifida fusca internally tangent beta-1, 4-glucanase |
| JP2015059071A (en) * | 2013-09-19 | 2015-03-30 | 秀夫 草桶 | Fertilizer containing chitin and/or chitosan or chitin and/or chitosan inclusion and culture of microorganisms having chitin and/or chitosan resolving ability, and production method thereof |
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| WO2020037866A1 (en) * | 2018-08-20 | 2020-02-27 | 广东药科大学 | Low molecular weight chitooligosaccharide and preparation method therefor |
| CN109438103A (en) * | 2019-01-10 | 2019-03-08 | 江苏心实肥业集团有限公司 | A kind of chitosan oligosaccharide crop nutrition spray fertilizer and preparation method thereof |
| CN113234774A (en) * | 2021-05-27 | 2021-08-10 | 厦门海洋职业技术学院 | Preparation method of chitosan oligosaccharide with high amino content |
| CN113583149A (en) * | 2021-08-13 | 2021-11-02 | 上海应用技术大学 | Production method for co-producing chitosan oligosaccharide and seafood flavor base material by taking crab shells as raw materials |
| CN115521960A (en) * | 2022-09-20 | 2022-12-27 | 山东海锋生物工程有限公司 | Production process for reducing non-enzymatic browning of chitosan oligosaccharide |
| CN115920015A (en) * | 2023-02-27 | 2023-04-07 | 广西师范大学 | Sulfate-crosslinked lysozyme hydrogel and preparation method thereof |
| CN115920015B (en) * | 2023-02-27 | 2024-04-26 | 广西师范大学 | Sulfate crosslinked lysozyme hydrogel and preparation method thereof |
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