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A method for screening marine microbial strains to degrade shrimp and crab shell powder to prepare chitin

A technology for marine microorganisms, shrimp and crab shell powder, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and microorganism determination/inspection, and can solve the problems of easy pollution, complicated preparation of culture medium, and cumbersome process.

Active Publication Date: 2019-10-29
浙江海富海洋生物科技有限公司
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  • Summary
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Because the chemical method used in the industrial preparation of chitin is easy to pollute the environment, change the product structure and introduce new impurities, while the existing biological method for preparing chitin, due to the high cost of preparing enzymes, difficulty in preservation and easy pollution, and the cultivation The preparation of the substrate is complicated, the process is cumbersome and many other problems. The present invention proposes a method for screening marine microbial strains to degrade shrimp and crab shell powder to prepare chitin. The technical scheme is as follows:

Method used

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  • A method for screening marine microbial strains to degrade shrimp and crab shell powder to prepare chitin
  • A method for screening marine microbial strains to degrade shrimp and crab shell powder to prepare chitin
  • A method for screening marine microbial strains to degrade shrimp and crab shell powder to prepare chitin

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preparation example Construction

[0036] Preparation of medium I: 5 grams of peptone, 1 gram of yeast extract, 0.01 gram of ferric phosphate, 20 grams of calcium carbonate, 15-20 grams of agar, 1000 ml of aged sea water, pH 7.6-7.8.

[0037] Screening of acid-producing microbial strains: Dilute seawater or sea mud taken from different sea areas with sterilized old seawater for 10 -1 、10 -2 、10 -3 、10 -4 times, spread samples of different dilutions on the plate of screening medium I, and then put the plate in a 25°C incubator for 2-3 days, select colonies with obvious transparent circles, and divide them three times and three divisions on 2216E Line, pick a single colony, which is the acid-producing microbial strain.

[0038] (2) Screening of protease-producing strains among acid-producing microbial strains:

[0039] Preparation of medium II: 5 grams of skimmed milk powder, 5 grams of peptone, 1 gram of yeast extract, 20 grams of agar powder, 1000 mL of aged sea water, pH 7.0-7.4.

[0040] Connect the sing...

Embodiment 1

[0053] Add a small amount of seawater and sea mud from different places into a 500mL triangular flask filled with 100mL of old seawater, add 5-10 sterilized glass beads in the bottle, and follow the gradient of 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 、10 -8 、10 -9 After dilution in sequence, spread on acid production identification plate respectively, acid production identification plate: 5 grams of peptone, 1 gram of yeast extract, 0.01 gram of ferric phosphate, 20 grams of calcium carbonate, 15-20 grams of agar, 1000 ml of aged sea water, pH7 .6-7.8, cultured in a 25°C incubator for 2-3 days. When colonies with transparent circles appear on the plate, pick 50 colonies with larger transparent circles and isolate single colonies on the 2216E plate using the three-section line method, and culture them in a 25°C incubator for 2-3 days. The single colonies screened on the 2216E plate were further streaked and purified 2-3 times to obtain acid-producing bacteria wi...

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Abstract

The invention discloses a method for screening marine microbial strains capable of degrading shrimp and crab shell powder and preparing chitin. The method is characterized by comprising the following steps: screening acid-producing microbial strains; screening protease-producing strains from the acid-producing microbial strains; screening the strains which can produce acid and produce protease, but cannot produce chitinase; screening the strains which can degrade the shrimp and crab shell powder and produce the chitin; identifying target strains 16S rDNA. Calcium carbonate, the protein and the chitin in the shrimp and crab shell powder are insoluble, so that the microbial strains which can degrade the shrimp and crab shell powder to produce the chitin are very difficult to screen by a single one-step operation; the target strains can be screened through several steps of a simple tablet microbial colony screening method; the method is simple and feasible, and is suitable for screening the strains which can degrade the shrimp and crab shell powder and produce the chitin from a marine environment.

Description

technical field [0001] The invention relates to a method for fermenting shrimp and crab shell powder to produce chitin, in particular to a method for preparing chitin by screening marine microbial strains to degrade shrimp and crab shell powder. Background technique [0002] Chitin, also known as chitin and chitin, is a structurally homopolysaccharide formed by the polymerization of N-acetylglucose through β-linkages. It is an off-white amorphous substance which becomes very hard when mixed with calcium salts. At present, most of chitin is produced by chemical methods. With the development of my country's aquaculture industry, more and more crab shells and shrimp shells are produced. At present, most of the shrimp and crab shells have no good treatment methods, which have caused a great degree of pollution to the environment. Studies have shown that the composition of shrimp and crab shells is composed of 20-30% chitin, 30-40% protein and minerals, of which the minerals ar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/02C12Q1/04C12Q1/689C12P19/04C12N1/20C12R1/63
CPCC12N1/02C12N1/20C12P19/04C12Q1/04C12Q1/689
Inventor 赵宏赵斌元孙玉英杨威刘茫茫
Owner 浙江海富海洋生物科技有限公司