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Aspergillus versicolor sd-3 and its application in the preparation of chitin deacetylase

A technology of Aspergillus versicolor, SD-3, applied to Aspergillus versicolor SD-3 and its application in the preparation of chitin deacetylase, which can solve the problems of low enzyme activity, long fermentation cycle, and high cost of medium, Achieve the effects of simple preparation process, easy cultivation, and simple nutritional requirements

Inactive Publication Date: 2016-06-01
ZHEJIANG SHUREN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a strain of chitin deacetylase producing bacteria - Aspergillus versicolor (Aspergillus versicolor) SD-3, and its application in the preparation of chitin deacetylase, better overcome the existing microbial fermentation to produce chitin The medium cost of deacetylase is high, the fermentation cycle is long and the enzyme activity is low. The production process has the advantages of low cost, simple process and high efficiency.

Method used

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  • Aspergillus versicolor sd-3 and its application in the preparation of chitin deacetylase
  • Aspergillus versicolor sd-3 and its application in the preparation of chitin deacetylase
  • Aspergillus versicolor sd-3 and its application in the preparation of chitin deacetylase

Examples

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Embodiment 1

[0027] Example 1: Enrichment, isolation and screening of chitin deacetylase-producing microorganisms

[0028] In order to obtain strains with high enzyme-producing activity, the chitin deacetylase-producing microorganisms were naturally enriched first. The method is: in the early summer season, bury crushed crab shells in the soil of flower beds, sprinkle water to keep the soil moist, collect soil samples after 2 months, and use them as the source of isolation of chitin deacetylase microorganisms.

[0029] Collect the naturally enriched soil, and then carry out artificial culture and enrichment of chitin deacetylase-producing microorganisms. The method is: add 45mL enrichment medium to a 250mL conical flask, add 5g of the above soil sample after sterilization, and place the conical flask at 28 ℃, 200r / min shaking culture for 5 days, take 5mL of the enriched culture medium and inoculate it into another Erlenmeyer flask containing 45mL of enriched medium, repeat the above cultur...

Embodiment 2

[0041] Embodiment 2: Mutation breeding of chitin deacetylase high-yielding strain

[0042] The wild Aspergillus versicolor CR-3 strain obtained by screening in Example 1 was mutated by ultraviolet irradiation, and the mutant strain with improved chitin deacetylase activity was screened.

[0043] Ultraviolet radiation mutagenesis method: After activating and culturing the slant strain of the strain CR-3, scrape the spores into a Erlenmeyer flask containing 50 mL of sterile saline, shake at room temperature for 30 minutes, put a cotton ball on the neck of the funnel to filter the spore suspension. Count the spores in the suspension with a hemocytometer under a microscope, and make appropriate multiple dilutions with sterile saline to control the number of spores at 1×10 8 A / mL or so. Under red light, take 2 mL of the above spore suspension and a sterile paper clip into a petri dish with a diameter of 6 cm, place the petri dish on a magnetic stirrer, and irradiate under a 20-min...

Embodiment 3

[0047] Embodiment 3: Method 1 for preparing chitin deacetylase by fermentation of Aspergillus versicolor SD-3

[0048] Using Aspergillus versicolor SD-3 as the bacterial species, the method for preparing chitin deacetylase at the scale of 50mL shake flask without seed expansion is as follows:

[0049] (1) The Aspergillus versicolor SD-3 strain preserved on the slant of the test tube was inoculated on the slant medium, and the slant was cultured in a biochemical incubator at 28° C. for 3 days. The composition and preparation method of the slant medium are the same as in Example 1.

[0050] (2) Scraping with an inoculation loop Step (1) After the activation culture, the Aspergillus versicolor SD-3 spores were inoculated into the enzyme-producing medium, and cultivated for 5 days at 28°C and 150r / min shaking conditions to obtain chitin deacetylase-containing Fermentation liquid; The described enzyme production medium consists of: sucrose 5g, yeast extract powder 2.5g, (NH 4 ) ...

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Abstract

The invention provides an Aspergillus versicolor SD-3 and its application in the preparation of chitin deacetylase. The Aspergillus versicolor SD-3 has simple nutrition requirements, has strong competitor pollution resistance, and is easy to culture; chitin deacetylase generated by the Aspergillus versicolor SD-3 is lyoenzyme, a crude chitin deacetylase liquid can be obtained after separating and removing the Aspergillus versicolor SD-3, and the crude chitin deacetylase liquid can be directly used, and is simple to prepare; the deacetylase produced by the Aspergillus versicolor SD-3 has high activity, and the vitality of the non-separated and non-purified crude deacetylase liquid reaches 142.3U / mL under optimal conditions; and a production technology of the chitin deacetylase does not use toxic harmful raw materials, so environmental protection and no pollution are realized.

Description

(1) Technical field [0001] The invention relates to a chitin deacetylase-producing Aspergillus versicolor SD-3 and its application in preparing the chitin deacetylase. (2) Background technology [0002] Chitin, also known as chitin, is a polysaccharide composed of N-acetylamino-D-glucose monomers linked by β-1,4-glucosidic bonds, widely present in many invertebrates (such as shrimp, The shell or cuticle of crabs, insects, etc.) is a natural polysaccharide second only to cellulose in nature. The molecular structure of chitin is difficult to dissolve in dilute acids and alkalis and general organic solvents due to its internal crystal structure and strong hydrogen bonding, so its application range is not wide. When the acetyl group in the chitin molecule is partially or completely removed, chitosan (chitosan) is formed. Due to its non-toxic, biodegradable, good biocompatibility and film-forming properties, it is widely used in medicine. , food, chemical industry, cosmetics an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N9/80C12R1/66
Inventor 陈虹张建芬柯薇活泼
Owner ZHEJIANG SHUREN UNIV
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