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Breeding method for cellulase high-yield bacteria

A cellulase and high-yield technology, applied in the field of cellulase bacteria, can solve the problems of reduced enzyme production capacity and easy degradation of strains

Inactive Publication Date: 2019-08-02
ANHUI WUYUE FOOD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many types of molds that produce cellulase, there is a serious defect in these molds: the strains are easy to degenerate, resulting in reduced enzyme production capacity

Method used

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  • Breeding method for cellulase high-yield bacteria

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Experimental program
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Effect test

Embodiment 1

[0029] 1) Trichoderma viride and Reesei were cultured in the first culture medium (containing 25mg / L nitrosoguanidine and 42mg / L diethyl sulfate) according to the colony ratio of 1:1.8 for 8h (can be 4- 8h), followed by 60 Co-γ rays (intensity of 60eV) were irradiated for 20 minutes to obtain mutant mixed mold strains;

[0030] 2) Cultivate Clostridium krusei thermocellulolyticus in the second medium (containing 12mg / L ethyl methanesulfonate, 20mg / L diethyl sulfate) for 4h (can be 1.5-4h), and then use Ultraviolet rays (provided by a 500w ultraviolet generator) were irradiated for 25 minutes to obtain mutant strains;

[0031] 3) The above-mentioned mutant mixed mold strains and mutant strains were cultured in MS medium according to the colony ratio of 1:1 to obtain cellulase high-producing bacteria.

Embodiment 2

[0033] 1) Trichoderma viride and Reesei were cultured in the first culture medium (containing 33mg / L nitrosoguanidine and 44mg / L diethyl sulfate) according to the colony ratio of 1:2 for 5h (can be 4- 8h), followed by 60 Co-γ rays (intensity of 80eV) were irradiated for 15 minutes to obtain mutant mixed mold strains;

[0034] 2) Cultivate Clostridium krusei thermocellulolyticus in the second medium (containing 16mg / L ethyl methanesulfonate, 22mg / L diethyl sulfate) for 2h (can be 1.5-4h), and then use Ultraviolet rays (provided by a 55w ultraviolet generator) were irradiated for 20 minutes to obtain mutant strains;

[0035] 3) The above-mentioned mutant mixed mold strains and mutant strains were cultured in MS medium according to the colony ratio of 1:1.3 to obtain cellulase high-producing bacteria.

Embodiment 3

[0037] 1) Trichoderma viride and Reesei were cultured in the first culture medium (containing 37mg / L nitrosoguanidine and 46mg / L diethyl sulfate) according to the colony ratio of 1:2.2 for 4h (can be 4- 8h), followed by 60 Co-γ rays (intensity of 100eV) were irradiated for 10 minutes to obtain mutant mixed mold strains;

[0038] 2) Cultivate Clostridium krusei thermocellulolyticus in the second medium (containing 18mg / L ethyl methanesulfonate, 25mg / L diethyl sulfate) for 1.5h (can be 1.5-4h), then Adopt ultraviolet light (provided by the ultraviolet generator of 600w) to irradiate 18min to obtain mutant strain;

[0039] 3) The above-mentioned mutant mixed mold strains and mutant strains were cultured in MS medium according to the colony ratio of 1:1.5 to obtain cellulase high-producing bacteria.

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Abstract

The invention discloses a breeding method for cellulase high-yield bacteria. The breeding method comprises the following steps: 1) culturing trichoderma viride and trichoderma reesei in a first medium, followed by 60 Co-gamma ray irradiation to obtain a mutant mixed mold; 2) culturing clostridium thermocellum in a second medium, followed by ultraviolet irradiation to obtain a mutant strain; 3) culturing the mutant mixed mold and the mutant strain in an MS medium to obtain a cellulase-producing strain, wherein the first medium contains nitrosoguanidine and diethyl sulfate; and the second mediumcontains ethyl methanesulfonate and diethyl sulfate. The cellulase high-yield bacteria selected by the method are resistant to degradation.

Description

technical field [0001] The invention relates to cellulase bacteria, in particular to a breeding method for cellulase high-yielding bacteria. Background technique [0002] Cellulase is a complex enzyme system composed of various hydrolytic enzymes. Many fungi in nature can secrete cellulase. Cellulase is a type of enzyme that biocatalyzes the decomposition of cellulose; a protein that can break down cellulose into oligosaccharides or monosaccharides. [0003] Cellulase is widely found in organisms in nature. Bacteria, fungi, and animals can produce cellulase. Cellulases generally used for production come from fungi, typically Trichoderma, Aspergillus and Penicillium. Although there are many types of molds that produce cellulase, there is a serious defect in these molds: the strains are easy to degenerate, resulting in reduced enzyme production ability. Contents of the invention [0004] The purpose of the present invention is to provide a method for breeding cellulase h...

Claims

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Application Information

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IPC IPC(8): C12N13/00C12N15/01C12R1/885C12R1/145
CPCC12N13/00C12N15/01
Inventor 许腾龙李娟朱荣荣何锋陆永明潘文娟
Owner ANHUI WUYUE FOOD
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