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Chitin deacetylase high-yielding strain and application thereof

A technology of tin deacetylase and crustacean is applied in the field of high-yielding strains of chitin deacetylase, which can solve the problems of failure to realize industrialization, long fermentation time, low enzyme activity, etc., and achieve broad industrial application prospects and rapid accumulation. Effect

Active Publication Date: 2018-09-18
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the production of chitin deacetylase by microbial fermentation has a series of problems such as long fermentation time and low enzyme activity, so the enzyme has not been industrialized so far

Method used

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  • Chitin deacetylase high-yielding strain and application thereof
  • Chitin deacetylase high-yielding strain and application thereof
  • Chitin deacetylase high-yielding strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The screening of embodiment 1 F6 bacterial strain

[0026] The soil samples collected in Shandong, Shenyang, Chengdu and other places were made into suspension liquid and spread on the enrichment medium (fine powder chitin 2.5g / L, dipotassium hydrogen phosphate 0.7g / L, magnesium sulfate 0.5g / L L, sodium chloride 0.1g / L), then the bacterial solution in the enrichment medium was diluted with sterile water and applied to the screening medium (colloidal chitin 2.5g / L, dipotassium hydrogen phosphate 0.7g / L, potassium dihydrogen phosphate 0.3g / L, magnesium sulfate 0.1g / L, p-nitroacetanilide 2.0g / L, agar 20g / L) at 30°C for 3 to 5 days. The obtained culture was transferred to a new solid plate, and the plate was streaked until a single colony appeared. After re-screening by the enzyme production test in the Erlenmeyer flask, a pure culture was finally obtained, numbered as F6 strain.

[0027] The 16S rDNA of the F6 strain was identified, and the phylogenetic tree was constru...

Embodiment 2

[0028] Example 2 Enzyme production by fermentation of Rhodococcus equi CGMCC No.14861

[0029] The Rhodococcus CGMCC No.14861 obtained in Example 1 was subjected to multi-batch fermentation culture.

[0030] (1) Seed cultivation

[0031] Culture conditions: stirring speed is 200rpm, temperature is 37°C, fermentation is 12h;

[0032] The seed medium is: peptone 10g / L, yeast extract powder 5g / L, sodium chloride 10g / L, the balance is water, pH6.0-7.0;

[0033] (2) Fermentation culture

[0034] Fermentation conditions: inoculum size 8%, stirring speed 200rpm, temperature 37°C;

[0035] Fermentation medium: yeast extract powder 5g / L, glucose 0.5g / L, magnesium sulfate 1.0g / L, potassium dihydrogen phosphate 0.3g / L, dipotassium hydrogen phosphate 1.0g / L, sodium chloride 0.5g / L, The balance is water, pH6.0-7.0;

[0036] After 30 hours of fermentation, the fermentation liquid was taken regularly to detect the activity of chitin deacetylase. Fermentation enzyme curve as image 3 A...

Embodiment 3

[0039] Example 3 Enzyme production by fermentation of Rhodococcus equi CGMCC No.14861

[0040] The Rhodococcus CGMCC No.14861 obtained in Example 1 was subjected to multi-batch fermentation culture.

[0041] (1) Seed cultivation

[0042] Culture conditions: stirring speed is 160rpm, temperature is 30°C, fermentation is 24h;

[0043] The seed medium is: peptone 5g / L, yeast extract powder 8g / L, sodium chloride 5g / L, the balance is water, pH6.0-7.0;

[0044] (2) Fermentation culture

[0045] Fermentation conditions: inoculum size 10%, stirring speed 160rpm, temperature 40°C;

[0046] Fermentation medium: yeast extract powder 10g / L, glucose 2.0g / L, magnesium sulfate 1.0g / L, potassium dihydrogen phosphate 0.3g / L, dipotassium hydrogen phosphate 1.0g / L, sodium chloride 2.0g / L, The balance is water, pH6.0-7.0;

[0047] When fermented to 6h, the enzyme activity of the fermentation broth reached 180.5U / mL.

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Abstract

The invention belongs to the biological technical field, and particularly relates to a chitin deacetylase high-yielding strain and an application thereof. The chitin deacetylase high-yielding strain is particularly rhodococcus usequi F6 and has the preservation number of CGMCC No.14861. The strain has quite fast chitin deacetylase producing speed, is rhodococcus usequi capable of producing chitindeacetylase and reported for the first time at present, is also a fastest strain of all strains for fermentation production of chitin deacetylase in the prior art, and is easy to culture on a large scale.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a chitin deacetylase high-yield bacterial strain and application thereof. Background technique: [0002] Chitin, also known as chitin, scientifically named (1,4)-2-acetylamino-2-deoxy-β-D-glucan, is an amino acid contained in natural organisms that is second only to cellulose Polysaccharides, mainly found in invertebrates, such as shrimp, insects, algae, fungi and yeast. However, chitin is insoluble in water, acid, alkali and organic solvents, so it has little commercial value, and the product chitosan after deacetylation is widely used in medicine, food, chemical industry, cosmetics and other industries. There are many problems in the chemical method currently used in the production of chitosan, such as long reaction time, high energy consumption, unstable product quality, and especially the discharge will cause huge environmental pollution. [0003] Chitin deacetylas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/80C12R1/01
CPCC12N1/20C12N9/80C12Y305/01041C12N1/205C12R2001/01
Inventor 马钦元王敏申雁冰夏梦雷毕心宇张子君杨阳
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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