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

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

Inactive Publication Date: 2014-10-22
ZHEJIANG SHUREN UNIV
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  • 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 ex

Method used

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  • Aspergillus versicolor SD-3 and its application in preparation of chitin deacetylase
  • Aspergillus versicolor SD-3 and its application in preparation of chitin deacetylase
  • Aspergillus versicolor SD-3 and its application in 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 higher enzyme-producing activity, chitin deacetylase-producing microorganisms were firstly enriched naturally. The method is as follows: in early summer, bury crushed crab shells in the soil of flower beds, sprinkle water to keep the soil moist, and collect soil samples here after 2 months as the isolation source of chitin deacetylase microorganisms.

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

Embodiment 2

[0041] Example 2: Mutation Breeding of Chitin Deacetylase High-Producing Strain

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

[0043] Ultraviolet irradiation mutagenesis method: After activating and culturing the slanted strains of strain CR-3, scrape the spores into a triangular flask containing 50 mL of sterile normal saline, shake at room temperature for 30 min, and place 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, make appropriate fold dilution with sterile normal saline, and control the number of spores at 1 × 10 8 pcs / mL or so. Under red light, take 2 mL of the above-mentioned 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 irradia...

Embodiment 3

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

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

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

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

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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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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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