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Highly effective chitosan degradation bacteria bacillus and its chitosan enzyme produced therefrom

A Bacillus, polycanase technology, applied in the direction of hydrolytic enzymes, microorganism-based methods, bacteria, etc., can solve the problems of products being a variety of mixtures, low productivity, etc.

Inactive Publication Date: 2010-03-10
THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have low productivity, and the products are difficult to overcome such as multiple mixtures.

Method used

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  • Highly effective chitosan degradation bacteria bacillus and its chitosan enzyme produced therefrom
  • Highly effective chitosan degradation bacteria bacillus and its chitosan enzyme produced therefrom
  • Highly effective chitosan degradation bacteria bacillus and its chitosan enzyme produced therefrom

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Screening of bacteria: preparation of chitosanase plate: screening required bacterial strains by the out-of-circle situation around different bacterial colonies on the plate containing 0.1% chitosan basic medium. And through 16S rDNA for preliminary identification.

[0011] Research on bacterial enzyme production: the screened bacteria were inserted into the basic culture solution containing 0.1% chitosan, and shake cultured overnight at 30°C. Concentrate the supernatant and break the cells, and add the concentrated supernatant and cell breakdown liquid into the McIlvaine buffer containing 0.01% chitosan, react at 37°C for 2 hours, and measure the content of reducing sugar in the reaction system by DNS method.

Embodiment 2

[0013] Research on the properties of enzyme production: through the growth of the strains under two different conditions of the basic culture medium with chitosan and no chitosan, and the enzymatic activity verification after the fermentation supernatant was concentrated as a crude enzyme, the results Show that the strain is non-induced enzyme production.

Embodiment 3

[0015] Determination of enzyme activity: the fermented supernatant is concentrated and used as crude enzyme, and the optimal reaction pH of the enzyme is PH6.5 through the determination of the amount of reducing sugar produced by enzymatic hydrolysis products of different pH gradients; and then reduced by different temperature gradients According to the efficiency of sugar production, the most suitable reaction temperature of this enzyme is 60℃.

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PUM

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Abstract

The invention discloses a Bacillus sp. to generate non-induced exocytotic chitosanase, which is characterized by the following: detecting enzyme activity of supernatant fermented by strain at 65 deg.c with pH value at 6. 0; displaying the final product as dichitosan and trichitosan through paper chromatography and mass spectrography.

Description

Technical field: [0001] The invention relates to a high-efficiency chitosan-degrading bacterium obtained through screening—bacillus, which secretes a chitosanase to degrade chitosan to obtain chitobiose and chitotriose. Background technique: [0002] Chitosan is a linear polymer of glucosamine linked by β-1,4 glycosidic bonds. It can be seen as a deacetylated derivative of chitin. It is widely present in the cell walls of zygomycetes and plant pathogens, and also exists in small amounts in the cell walls of green algae, fungi, and some human pathogens. Chitosanase (EC3.2.1.99) can degrade the β-1,4 glycosidic bond of chitosan, and can be divided into endonuclease and exonuclease. Chitosan endonuclease can randomly cut polysaccharide chains (n≥4) to obtain chitosan oligosaccharides with different molecular weights. Many microorganisms can secrete chitosanase, which degrades chitosan to provide carbon and nitrogen sources needed for growth. [0003] Chitosan mainly refers ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P19/00C12P19/12C12P19/14C12N9/24C12R1/07
Inventor 肖湘王凤平汤熙翔游志勇
Owner THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION
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