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Chitosanase producing strain and chitosan production method by using the same

A technology for producing chitosanase and strains, which is applied in the field of bioengineering, can solve the problems of not fundamentally reducing the production cost of chitosan oligosaccharide, and the process of breaking cells is complicated and reduced.

Active Publication Date: 2013-01-02
CHAMBROAD CHEM IND RES INST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above two types of chitosan enzyme preparations are all liquid enzymes. Although they are convenient to use, they are not conducive to storage and transportation, which limits the application of the enzyme to a certain extent.
Chinese patent CN 101397752A adopts genetic engineering means to construct chitosanase high-efficiency expression strains. Its chitosanase expression carrier is Escherichia coli, and the fermented liquid enzyme activity is 350U / ml. Intracellular enzymes, bacterial cell disruption process is relatively complicated, which affects the reduction of production costs
[0004] The bacterial strains reported in the above several documents have much higher activity of chitosanase than those reported in other documents, but there are still various defects and deficiencies, which are mainly manifested in the fact that the enzyme activity is still high from the perspective of enzyme preparation production and utilization. Low, high production costs, and liquid enzyme preparations are not conducive to storage and transportation, etc., did not fundamentally reduce the production cost of chitosan oligosaccharides, limiting its large-scale application

Method used

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  • Chitosanase producing strain and chitosan production method by using the same
  • Chitosanase producing strain and chitosan production method by using the same
  • Chitosanase producing strain and chitosan production method by using the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] liquid seed culture

[0070] The composition of seed medium (w / w) is: beef extract 2%, yeast powder 0.5%, glucose 2%, ammonium sulfate 0.5%, soybean meal 3%, corn flour 3%, and the balance is water; initial pH value is natural; 121 ℃, sterilize at 0.15Mpa for 20min.

[0071] Seed culture conditions: under aseptic conditions, rinse the activated CGMCC No.6129 test-tube slanted strains into a conical flask containing sterilized liquid medium with 10 mL of sterilized distilled water, and inoculate one inoculated with one test-tube strain Erlenmeyer flask, culture temperature 33~35℃, shaker speed 150r / min, culture time 12~18h;

Embodiment 2

[0073] solid fermentation

[0074] Composition of solid fermentation medium (w / w): 70% bran, 15% soybean meal, 13% corn meal, 2% ammonium sulfate; the ratio of material to water is 1:1.1; the initial pH value is natural; the culture temperature is 33 °C, and the fermentation The time is 48h; every 15h, the song is turned over.

[0075] After the fermentation is completed, it is dried at 55°C, pulverized, and passed through an 80-mesh sieve, which is the crude chitosanase enzyme. Samples were taken and the enzyme activity was detected by DNS method, and the average enzyme activity was 2547U / g.

Embodiment 3

[0077] solid fermentation

[0078] Composition of solid fermentation medium (w / w): 80% bran, 10% soybean meal, 7% corn meal, 3% ammonium sulfate; the ratio of material to water is 1:1.15; the initial pH value is natural; the cultivation temperature is 33~35℃ , the fermentation time is 48h; it is turned every 15h.

[0079] After the fermentation was completed, it was dried at 50°C, crushed, and sieved to obtain the crude chitosanase enzyme. The chitosanase activity was sampled and detected, and the average enzyme activity was 2732 U / g.

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Abstract

The invention belongs to the technical field of biological engineering, and provides a chitosanase high-yield producing strain, a liquid and solid fermentation method for preparing chitosanase with high activity by using the strain, and chitosan oligosaccharide production method by using the chitosanase. The chitosanase producing strain in the invention is identified as a strain of Bacilluscereus with a strain code of JBSH-003 by 16SrDNA identification, and has a preservation number of CGMCCNo.6129 in China General microbiological culture collection center. The strain can be applied to the production of chitosan to obtain chitosan oligosaccharide with high content, molecular weight range of 1500Dalton-2500Dalton, and relatively concentrated molecular weight range.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and provides a chitosanase-producing microbial strain, in particular to a Bacillus cereus strain, and a method for using the bacteria to produce chitosanase and using the enzyme to produce chitosan oligosaccharide . Background technique [0002] Chitosan oligosaccharide is a low molecular weight product of chitosan, mostly refers to low molecular weight chitosan with a degree of polymerization between 2 and 12 and an average molecular weight between 300 and 3000 Daltons. Because chitosan oligosaccharide has greater biological application value than chitosan, it has increasingly become a research hotspot of domestic and foreign scholars. At present, there are three main ways of chitosan oligomerization, physical method (mainly refers to ultrasonic method), chemical method (mainly chemical synthesis method and chemical degradation method) and biological enzyme method. In the past 20 years,...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/14C12R1/085
Inventor 徐泽平马韵升史庆苓杨传伦张心青王秀芝虞凤慧高洪奎
Owner CHAMBROAD CHEM IND RES INST CO LTD
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