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Method for breeding high-producing strains through acid protease by means of ultraviolet mutation

A technology of acid protease and high-yield strains, applied in biochemical equipment and methods, enzymes, electric/wave energy treatment enzymes, etc., can solve the problems of low efficiency and small increase in yield, and achieve simple equipment, convenient operation, Inexpensive effect

Inactive Publication Date: 2016-07-20
ZHEJIANG UNIV OF TECH
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Problems solved by technology

The currently reported strain breeding methods are mainly a combination of traditional chemical mutagenesis methods (nitrosoguanidine, diethyl sulfate, ethyl methanesulfonate, etc.) and physical mutagenesis methods. low, and the rate of increase in output is not large

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  • Method for breeding high-producing strains through acid protease by means of ultraviolet mutation
  • Method for breeding high-producing strains through acid protease by means of ultraviolet mutation

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Embodiment Construction

[0019] The acid protease high-yielding bacterial strain selection method that adopts ultraviolet mutagenesis in the present embodiment may further comprise the steps:

[0020] a) Preparation of spore suspension: Take the plate spores cultured at 35°C for 8 days in a constant temperature incubator, elute the plate spores with sterile distilled water, place them in a sterile triangular flask filled with glass beads, and heat them at 250-400r / min Vibrate on a special vibrating device for 6-10 hours to activate and disperse the spores. The spore dispersion rate is required to be above 95%. Observe and count the spores under an optical microscope, and then dilute the spore suspension to 126 spores / mL with sterile distilled water to obtain The spore suspension is ready for use; the shaking device is a floor-standing large-capacity constant temperature shaking shaker, the shaking speed is 250r / min, and the shaking time is 8h;

[0021] b) Ultraviolet mutagenesis: Take 5mL of the spore...

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Abstract

The invention discloses a method for breeding high-producing strains through acid protease by means of ultraviolet mutation.The method comprises the following steps of a, preparation of spore suspension liquid, wherein tablet spores cultured in a constant temperature incubator are taken, shaken and then diluted with sterile distilled water, and the spore suspension liquid is obtained for use; b, ultraviolet mutation, wherein the spore suspension liquid is taken and put under an ultraviolet lamp for irradiation, culture is carried out under a dark condition, and finally the fatality rate is calculated; c, strain screening, wherein c1, based on strains with the fatality rate between 70% and 80% in the step b, variant strains with the HC value higher than the mean value are picked out as preliminarily screened strains for secondary screening; c2, a conical flask containing a potato culture medium is inoculated with the strains obtained in the step c1, and after culture in a shaker, the enzymatic activity of acid protease is measured through a foline-phenol method; the mutated variant strains with the enzymatic activity higher than the mean value are selected for six times of continuous passage, and the high-producing strains are obtained.

Description

technical field [0001] The invention belongs to the technical field of strain breeding, and in particular relates to a method for breeding acid protease high-yield strains by ultraviolet mutagenesis. Background technique [0002] The strains isolated and screened by natural breeding methods in nature have the disadvantages of low strain vigor and low resistance to physical and chemical factors, which cannot meet the needs of industrial production. Therefore, mutation breeding has become the only way to solve this problem. , and has become the most commonly used method for industrial microbial breeding. Mutation breeding is the process of artificially treating microorganisms to induce mutations and using reasonable screening procedures and methods to select and breed excellent strains suitable for human needs. It can greatly increase the mutation rate of microorganisms and is a very important breeding method. . The main steps of mutation breeding are divided into two parts:...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N13/00C12N1/02
CPCC12N13/00C12N1/02
Inventor 何荣军王茜孙培龙
Owner ZHEJIANG UNIV OF TECH
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