Strain for producing chitinase and application of strain

A chitinase and chitin technology, applied in the direction of enzymes, bacteria, enzymes, etc., can solve the problems of microbial diversity and complex composition, and achieve the effects of easy cultivation and preservation, good growth and broad application prospects

Active Publication Date: 2019-02-15
CHANGSHU INSTITUTE OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The microbial diversity and composition of hemp shrimp paste are very complex, which is suitable for the screening of chitinase-producing strains. At the same time, hemp shrimp is rich in chitin and low in price. It is suitable as a substrate for chitin decomposition products and has strong industrial applications Foreground, there is currently no report on screening chitinase-producing strains from hemp shrimp paste and using hemp shrimp to produce chitooligosaccharides

Method used

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  • Strain for producing chitinase and application of strain
  • Strain for producing chitinase and application of strain
  • Strain for producing chitinase and application of strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] This example illustrates that Streptomyces amylase ( Streptomyces diastaticus ) Screening, purification and identification methods of CS1801.

[0039] The screened sample was hemp shrimp paste from Lianyungang Haiwa Food Co., Ltd. Weighed 25g of shrimp paste and 225mL of normal saline to prepare a bacterial suspension, and diluted their concentrations to 10 -1 、10 -2 、10 -3 and 10 -4 times. The bacterial suspension stock solution, 10 -1 Doubly diluted bacterial solution, 10 -2 Doubly diluted bacterial solution, 10 -3 Doubly diluted bacterial solution and 10 -4 The double-diluted bacterial solution was spread on the primary screening medium, and after growing at 37°C for 1-7 days, a single colony with good growth was picked and separated by streaking on the primary screening medium. Pick a single colony with an obvious transparent circle around it produced on the primary screening medium and inoculate it into the liquid medium, and culture it on a shaker at 37°C ...

Embodiment 2

[0046] This example specifically illustrates the application of strain CS1801 in the production of chitooligosaccharides by colloidal chitin fermentation.

[0047] (1) Inoculate the strain CS1801 into PDA liquid medium, and culture with shaking at 30°C for 2-3 days;

[0048] (2) Inoculate the strain cultivated in step (1) into the fermentation medium, and culture with shaking at 30°C for 7 days. The fermentation broth was centrifuged, the precipitate was discarded, and the supernatant was taken. The chitin enzyme activity was determined to be 117.4 U / L, and the chitooligosaccharide content was 1.18 mol / L.

[0049] The composition of the fermentation medium is the same as that of the aforementioned rescreening medium.

[0050] The assay method of chitinase activity is as follows: centrifuge the fermentation broth at 3000 r for 10 min, and take the supernatant as the sample to be tested. Add the sample to be tested and horseradish peroxidase (HRP)-labeled detection antibody in...

Embodiment 3

[0052] This example specifically illustrates the types of products produced by strain CS1801 using colloidal chitin to ferment chitooligosaccharides.

[0053] (1) Inoculate the strain CS1801 into PDA liquid medium, and culture with shaking at 30°C for 2-3 days;

[0054] (2) Inoculate the strain cultivated in step (1) into the fermentation medium, and culture with shaking at 30°C for 7 days. The fermentation broth was centrifuged, the precipitate was discarded, and the supernatant was taken.

[0055] The composition of the fermentation medium is the same as that of the aforementioned rescreening medium.

[0056] Using ultra-high performance liquid chromatography-mass spectrometry (UPLC-Q-TOF-MS) technology to analyze the components in the fermentation broth, it was determined that chitin was decomposed into chitobiose, chitotriose, and chitotetraose under the action of Streptomyces CS1801 , chitopentaose and chitosan hexaose. Ultra-high performance liquid chromatography see ...

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Abstract

The invention relates to a strain for producing chitinase and application of the strain. The strain is classified and named as (Streptomyces diastaticus) CS1801 with a preservation number of CCTCC NO:M2018263. The Streptomyces diastaticus CS1801 comes from naturally-fermented metapenaeus affinis paste. By fermenting the metapenaeus affinis, the enzyme activity of the chitinase is as high as 57.3U / L and the content of chitooligosaccharides is 0.58mol / L. The invention provides a novel method for producing the chitooligosaccharides and has a good application prospect.

Description

technical field [0001] The invention relates to a chitinase-producing bacterial strain and its application in fermenting hemp shrimp to produce chitooligosaccharides, belonging to the technical field of industrial microorganisms. Background technique [0002] Chitin, also known as chitin, is a polymer of N-acetyl-D-glucosamine linked by β-1,4 glycosidic bonds. It exists widely in nature, and its reserves are second only to cellulose. The water insolubility of chitin greatly limits the scope of application, but it can be decomposed into water-soluble chitooligosaccharides or chitooligosaccharides (COS) under the action of chitinase. Its decomposition products have various functions such as antibacterial, anti-oxidation, and promoting the growth and development of animals and plants. A large number of studies have found that chitosan and chitosan oligosaccharides have significant anti-neuroinflammation and anti-oxidation effects, and may be widely used in the treatment of Alz...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/42C12P19/14C12P19/00C12P19/04C12R1/465
CPCC12N9/2442C12P19/00C12P19/04C12P19/14C12Y302/01014C12R2001/465C12N1/205C12N1/20
Inventor 王立梅徐田甜齐斌
Owner CHANGSHU INSTITUTE OF TECHNOLOGY
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