CN104045738B - A kind of composite fermentation enzymolysis is prepared the method for chitosan - Google Patents
A kind of composite fermentation enzymolysis is prepared the method for chitosan Download PDFInfo
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- CN104045738B CN104045738B CN201310450580.XA CN201310450580A CN104045738B CN 104045738 B CN104045738 B CN 104045738B CN 201310450580 A CN201310450580 A CN 201310450580A CN 104045738 B CN104045738 B CN 104045738B
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Abstract
The invention discloses a kind of composite fermentation enzymolysis and prepare the method for chitosan, comprise the steps: dry to crab shell or shrimp shell, pulverizing; Fermentation; Enzymolysis decalcification and de-albumen; De-acetyl, washing and drying are pulverized and be get final product. Enzymolysis process of the present invention is compared with prior art system, and decalcification and de-albumen are realized in same step, greatly enhance productivity, and need not use strong acid, consume energy low, pollution-free, environmental protection more. Free calcium solution after enzymolysis can be prepared into organic calcium preparation after reclaiming, and all like this products can both be used appropriately and produce significant economic worth.
Description
Technical field
The present invention relates to chitosan preparation method, belong to manufacture field.
Background technology
Chitin is a kind of natural nitrogenous polysaccharide material, extensively in the shell or cuticula of existence and invertebrate, especially in the shell of shrimp crab class. Chitin is easy to obtain, renewable, is to be only second to cellulosic natural homopolymer, but because intramolecule has very strong hydrogen bond action, dissolubility is low, and range of application is restricted.
Generating chitosan after chitin deacetylase. The preparation of shitosan at present adopts chemical method and the de-acetyl of enzymatic isolation method more. In chemical method deacetylation, the factor of the de-acetyl of impact is a lot, but major influence factors comprises: concentration of lye, reaction temperature and time. In general, improve reaction temperature and prolongation reaction time and all can improve deacetylation, but can cause the degraded of chitin main chain to increase, thereby affect viscosity and the molecular weight of product. Pass into the degraded that nitrogen can slow down main chain to a certain extent. The concentration of lye using due to chemical method is high, and the reaction time is long, unstable product quality, and environmental pollution is serious. The product that traditional chemical method is prepared after shitosan decalcification and de-albumen goes out of use, and cannot recycle, and causes waste.
Therefore, a kind of method that find that strong acid and highly basic consumption are few, pollution-free, decalcification and de-protein product can be recovered utilization will have splendid market prospects, and this also becomes those skilled in the art and is badly in need of the technical problem solving.
Summary of the invention
The object of the present invention is to provide a kind of new chitosan preparation method, described method can be compared with more energy-conserving and environment-protective of prior art.
Another object of the present invention is to provide a kind of safe chitosan preparation method, and the chitosan that described method makes is residual without strong acid or highly basic, and product is safer.
Another object of the present invention is to provide the chitosan preparation method that a kind of intermediate product can be recycled, to obtain higher economic worth.
Another object of the present invention is to provide a kind of chitosan with the market competitiveness, and described product is not because the improvement of technique increases production cost.
For achieving the above object, the present invention adopts following technical proposal to realize:
Dry, ultramicro grinding, fermentation, enzymolysis decalcification and albumen, filtration; Take off acetyl, washing and drying and get final product.
Crab shell or shrimp shell before the present invention adopts preparation method to decalcification carry out pretreatment, and then carry out enzymolysis decalcification and de-albumen.
For strengthening decalcification and de-albumen effect, the present invention is preferably broken to 0.5～1.0mm particle by crab shell or shrimp shell meal, the too small production cost that increases of particle, and the excessive effect of particle is poor. Inventor, through test of many times and experience, finally determines that 0.5～1.0mm granulometric range reaches the best cost performance of decalcification and cost.
Inventor provides great many of experiments, unexpected find various lactobacillus ferment according to certain proportioning combination after decalcification effect better, and it is better to adopt bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and three kinds of lactic acid bacterias of Lactobacillus helveticus (L.helviticus) to carry out ferment effect; Adopt these three kinds of lactic acid bacteria proportioning after fermentation effects best, in the time that three kinds of lactic acid bacterias combine with weight ratio 2:6:5, after the more single lactobacillus-fermented of free calcium content, improve 10%～20%. Preferably, lactic acid bacteria accounts for crab shell or shrimp husk as raw material weight ratio is 0.02～0.06%.
The present invention adopts the method for high-temperature sterilization to reach the object of sterilizing, also removes the lactic acid bacteria in enzymolysis liquid simultaneously.
Enzymolysis of the present invention adopts complex enzyme hydrolysis method, and after screening, final discovery is carried out enzymolysis with papain and bromelain and had better effect compared with other protease. Certainly, select other protease to carry out enzymolysis and also can realize object of the present invention. In technique of the present invention, best enzymolysis scheme is in the raw material after broken, to add the water of 4-8 times of w/v to make feed liquid on crab shell or shrimp shell meal, adds lactobacillus-fermented 36h, then 70～100 DEG C, and 20min; Zymotic fluid adds the compound protease of bromelain and papain weight ratio 1:2～5, and adding citric acid is adjusted pH value to 5.5～6.5, and 40～60 DEG C are stirred 1～2h; Separating liquid, residue repeats to extract again.
It is 0.05～0.2% for good that enzymolysis shell material compound protease used accounts for shell material weight ratio.
Enzymolysis, except having Deproteinated effect, also has the effect of decalcification. Enzymolysis can dissociate calcium constituent in the product after fermentation out, reaches decalcification effect.
After the de-albumen of enzymolysis, filter, filtrate recovery is dried to obtain organic calcium. In residue, add the sodium hydroxide solution of 40-60%, 60-90 DEG C, 1.5-4h, is washed to neutrality, is drying to obtain chitosan.
After enzymolysis decalcification and de-albumen, the calcium ion in raw material, other organic substance of protein are removed.
Chitosan deacetylation of the present invention is up to more than 96%.
The various formulations that in product of the present invention and any or more than one pharmacies, auxiliary material is mixed as starch, dextrin, lactose, microcrystalline cellulose, HPMC, polyethylene glycol, dolomol, superfine silica gel powder, xylitol, lactitol, glucose, glycine, sweet mellow wine, glycine etc., for example, can be made into tablet, sustained release tablets, dripping pill, granule, capsule, fine granule. Preferred dosage form is capsule or granule.
Enzymolysis process of the present invention is compared with prior art system, and decalcification and de-albumen are realized in same step, greatly enhance productivity, and need not use strong acid, consume energy low, pollution-free, environmental protection more. Free calcium solution after enzymolysis can be prepared into organic calcium preparation after reclaiming, and all like this products can both be used appropriately and produce significant economic worth.
Detailed description of the invention
Embodiment 1
To 0.5mm particle, add the water of 4 times of w/vs to make feed liquid crab shell or shrimp shell drying and crushing, add bifidobacterium lactis: lactobacillus bulgaricus: the lactic acid bacteria of Lactobacillus helveticus=2:6:5, it is 0.02% that lactic acid bacteria accounts for raw material weight ratio, fermentation 36h, then 70～100 DEG C, 20min; Zymotic fluid adds the compound protease of bromelain and papain weight ratio 1:2, and it is 0.05% that protease accounts for shell material weight ratio, and adding citric acid is adjusted pH value to 5.5, and 40 DEG C are stirred 1h; Separating liquid, residue repeats to extract again, filters, and abandons filtrate, leaves and takes residue, adds 40% sodium hydroxide solution, and 60 DEG C, 1.5h, is washed to neutrality, is drying to obtain chitosan.
Embodiment 2
By crab shell or shrimp shell drying and crushing to 1.0mm particle, add the water of 8 times of w/vs to make feed liquid, add bifidobacterium lactis: lactobacillus bulgaricus: the lactic acid bacteria of Lactobacillus helveticus=2:6:5, it is 0.06% that lactic acid bacteria accounts for raw material weight ratio, zymotic fluid adds the compound protease of bromelain and papain weight ratio 1:5, it is 0.2% that protease accounts for shell material weight ratio, and adding citric acid is adjusted pH value to 6.5, and 60 DEG C are stirred 2h; Separating liquid, residue repeats to extract again, filters, and abandons filtrate, leaves and takes residue, adds 60% sodium hydroxide solution, and 90 DEG C, 4h, is washed to neutrality, is drying to obtain chitosan.
Embodiment 3
By crab shell or shrimp shell drying and crushing to 0.5mm particle, add the water of 8 times of w/vs to make feed liquid, add bifidobacterium lactis: lactobacillus bulgaricus: the lactic acid bacteria of Lactobacillus helveticus=2:6:5, it is 0.04% that lactic acid bacteria accounts for raw material weight ratio, zymotic fluid adds the compound protease of bromelain and papain weight ratio 1:5, it is 0.1% that protease accounts for shell material weight ratio, and adding citric acid is adjusted pH value to 6.5, and 60 DEG C are stirred 2h; Separating liquid, residue repeats to extract again, filters, and abandons filtrate, leaves and takes residue, adds 60% sodium hydroxide solution, and 90 DEG C, 4h, is washed to neutrality, is drying to obtain chitosan.
Embodiment 4
By crab shell or shrimp shell drying and crushing to 1.0mm particle, add the water of 4 times of w/vs to make feed liquid, add bifidobacterium lactis: lactobacillus bulgaricus: the lactic acid bacteria of Lactobacillus helveticus=2:6:5, it is 0.05% that lactic acid bacteria accounts for raw material weight ratio, zymotic fluid adds the compound protease of bromelain and papain weight ratio 1:2, it is 0.15% that protease accounts for shell material weight ratio, and adding citric acid is adjusted pH value to 6, and 40 DEG C are stirred 2h; Separating liquid, residue repeats to extract again, filters, and abandons filtrate, leaves and takes residue, adds 60% sodium hydroxide solution, and 60 DEG C, 1.5h, is washed to neutrality, is drying to obtain chitosan.
Embodiment 5
By crab shell or shrimp shell drying and crushing to 0.8mm particle, add the water of 6 times of w/vs to make feed liquid, add bifidobacterium lactis: lactobacillus bulgaricus: the lactic acid bacteria of Lactobacillus helveticus=2:6:5, it is 0.03% that lactic acid bacteria accounts for raw material weight ratio, zymotic fluid adds the compound protease of bromelain and papain weight ratio 1:3, it is 0.12% that protease accounts for shell material weight ratio, and adding citric acid is adjusted pH value to 5.5, and 50 DEG C are stirred 2h; Separating liquid, residue repeats to extract again, filters, and abandons filtrate, leaves and takes residue, adds 50% sodium hydroxide solution, and 80 DEG C, 3h, is washed to neutrality, is drying to obtain chitosan.
Embodiment 6
By crab shell or shrimp shell drying and crushing to 0.9mm particle, add the water of 5 times of w/vs to make feed liquid, add bifidobacterium lactis: lactobacillus bulgaricus: the lactic acid bacteria of Lactobacillus helveticus=2:6:5, it is 0.06% that lactic acid bacteria accounts for raw material weight ratio, zymotic fluid adds the compound protease of bromelain and papain weight ratio 1:2.5, it is 0.18% that protease accounts for shell material weight ratio, and adding citric acid is adjusted pH value to 6.5, and 55 DEG C are stirred 1h; Separating liquid, residue repeats to extract again, filters, and abandons filtrate, leaves and takes residue, adds 55% sodium hydroxide solution, and 70 DEG C, 2.5h, is washed to neutrality, is drying to obtain chitosan.
Embodiment 7
By crab shell or shrimp shell drying and crushing to 0.6mm particle, add the water of 7 times of w/vs to make feed liquid, add bifidobacterium lactis: lactobacillus bulgaricus: the lactic acid bacteria of Lactobacillus helveticus=2:6:5, it is 0.05% that lactic acid bacteria accounts for raw material weight ratio, zymotic fluid adds the compound protease of bromelain and papain weight ratio 1:4, it is 0.08% that protease accounts for shell material weight ratio, and adding citric acid is adjusted pH value to 5.5, and 45 DEG C are stirred 1.5h; Separating liquid, residue repeats to extract again, filters, and abandons filtrate, leaves and takes residue, adds 48% sodium hydroxide solution, and 75 DEG C, 3h, is washed to neutrality, is drying to obtain chitosan.
Claims (2)
1. composite fermentation enzymolysis is prepared a method for chitosan, comprises the steps:
(1) crab shell or shrimp shell are dried, are crushed to 0.5～1.0mm granular size;
(2) lactobacillus-fermented, described lactic acid bacteria is bifidobacterium lactis, lactobacillus bulgaricus and Lactobacillus helveticus compound bacteria, lactic acid bacteria accounts for crab shell or shrimp husk as raw material weight ratio is 0.02～0.06%, and bifidobacterium lactis, lactobacillus bulgaricus, Lactobacillus helveticus three's weight ratio is 2:6:5;
(3) protease hydrolyzed decalcification and de-albumen: the compound protease that adds bromelain and papain weight ratio 1:2～5 in zymotic fluid, it is 0.05～0.2% that described compound protease accounts for shell material weight ratio, adding citric acid adjust pH to 5.5～6.5,40～60 DEG C are stirred 1～2h;
(4) de-acetyl, washing and drying are pulverized and be get final product.
2. preparation method according to claim 1, is characterized in that, described de-acetyl is the sodium hydroxide solution that adds 40-60% in residue, 60-90 DEG C, and 1.5-4h, is washed to neutrality, is drying to obtain.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104480228A (en) * | 2014-11-25 | 2015-04-01 | 荣成宏业海洋科技有限公司 | Method for preparing glucosamine by utilizing shrimp and crab shells |
| CN105154092B (en) * | 2015-08-21 | 2019-02-19 | 北京雷力海洋生物新产业股份有限公司 | A kind of soil heavy metal passivant and preparation method thereof |
| CN108440799A (en) * | 2018-04-20 | 2018-08-24 | 常州市蒽盗钟情生物科技有限公司 | A kind of preparation method of modification of chitosan base bionic fish bait |
| CN109160958A (en) * | 2018-08-07 | 2019-01-08 | 东山县吉兴水产加工有限公司 | A kind of technique of the chitin extraction from shrimp crab |
| CN110627922A (en) * | 2019-10-31 | 2019-12-31 | 山东花物堂生物科技有限公司 | Extraction process of chitosan in crab shells |
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| CN101144097A (en) * | 2007-09-18 | 2008-03-19 | 重庆百奥帝克微生态科技有限公司 | Method for preparing chitin and its chitosan and chitosan oligosaccharide |
| CN101869171A (en) * | 2010-05-18 | 2010-10-27 | 华南农业大学 | Method for extracting proteins and chitin from shrimp heads and shrimp shells by lactobacillus plantarum |
| MX2011006064A (en) * | 2011-06-08 | 2012-06-21 | Univ Autonoma Metropolitana | Bioreactor for obtaining chitin and astaxanthin by using lactobacillus and proteases resulting from shrimp wastes. |
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| BRPI0804835A2 (en) * | 2008-11-06 | 2011-05-24 | Quantas Biotecnologia S.A. | continuous industrial process for the production of polysaccharide in the form of concentrated water-based broth |
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| CN101144097A (en) * | 2007-09-18 | 2008-03-19 | 重庆百奥帝克微生态科技有限公司 | Method for preparing chitin and its chitosan and chitosan oligosaccharide |
| CN101869171A (en) * | 2010-05-18 | 2010-10-27 | 华南农业大学 | Method for extracting proteins and chitin from shrimp heads and shrimp shells by lactobacillus plantarum |
| MX2011006064A (en) * | 2011-06-08 | 2012-06-21 | Univ Autonoma Metropolitana | Bioreactor for obtaining chitin and astaxanthin by using lactobacillus and proteases resulting from shrimp wastes. |
Non-Patent Citations (2)
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| "Chitin and chitosan preparation from shrimp shells using optimized enzymatic deproteinization";Islem Younes等;《Process Biochemistry》;20120901;第47卷;第2032-2039页 * |
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