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
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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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