A kind of artificially induced Geobacillus thermoglucosidase and its application in cellulose degradation

A thermoglucosidase, Geobacillus technology, applied in the field of microorganisms, can solve the problem that P.thermoglucosidasius has not been reported in cellulose degradation, and achieve the effect of high enzyme concentration

Active Publication Date: 2022-08-09
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The ability of P.thermoglucosidasius to degrade cellulose has not been reported so far

Method used

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  • A kind of artificially induced Geobacillus thermoglucosidase and its application in cellulose degradation
  • A kind of artificially induced Geobacillus thermoglucosidase and its application in cellulose degradation
  • A kind of artificially induced Geobacillus thermoglucosidase and its application in cellulose degradation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Isolation, purification and identification of high temperature cellulose degrading bacteria

[0029]Pig carcass and straw compost samples were separated and purified in beef extract peptone medium, enriched in sodium carboxymethyl cellulose basal medium, and selected strains were spotted on cellulose Congo red medium, cultured at 60°C for 72 hours, and after primary screening, A total of 27 single strains were obtained from the sodium carboxymethyl cellulose medium plate, and then re-screened, the cellulase activity of each strain was determined, and a strain with the highest enzyme activity was finally selected, which was identified as Geobacillus by sequencing thermodenitrificansCMC-4, after high temperature acclimation and ultraviolet mutagenesis, a strain with high cellulase production was obtained. The strain has been sent to the China Center for Type Culture Collection for preservation on July 16, 2020, preservation number: CCTCC NO: M2020324, taxonomic name: Para...

Embodiment 2

[0033] Thermoglucosidase Geobacillus Parageobacillus thermoglucosidasius BGSC95A1 Cellulase Activity Determination:

[0034] The strain Parageobacillus thermoglucosidasius BGSC95A1 and its original strain Geo bacillus thermodenitrificansCMC-4 were respectively inoculated into the liquid fermentation medium with 2% inoculum, and cultured at 60°C and 150r / min for 72h, and the strain fermentation broth was centrifuged at 8000r / min for 30min to remove bacteria. body and medium, and the supernatant is the crude enzyme solution.

[0035] The formula of the liquid fermentation medium: beef extract 10g, peptone 10g, yeast extract 10g, sodium chloride 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.2g, distilled water to make up to 1000mL, natural pH. Sterilize at 121°C for 20min.

[0036] Extract the crude enzyme solution, measure its carboxymethyl cellulase enzyme activity, and the detection method of the enzyme activity (that is, the enzyme concentration) is as follows: ...

Embodiment 3

[0049] Effects of different carbon sources on the enzyme production of Geobacillus thermoglucosidase BGSC95A1:

[0050] 1) 1% carboxymethyl cellulose sodium (CMC-Na), 1% microcrystalline cellulose (micro-cellulose), 1% soluble starch (starch), 1% glucose (glucose), 1% straw (straw) respectively ) was the carbon source, and the pH was adjusted to 7.2. The activated Geobacillus thermoglucosidase BGSC95A1 bacterial broth (ie, CMC-4-1) was inoculated into a 250 mL conical flask containing 50 mL of liquid fermentation medium at an inoculum of 2%. After culturing at 55°C and 200r / min shaker for 72h, the strain fermentation broth was centrifuged at 8000r / min for 30min to remove the bacteria and medium, and the supernatant was the crude enzyme solution; each group was repeated 3 times without adding any carbon source , peptone as nitrogen source is CK group; cellulase activity (method of Example 2) is measured, the enzyme activity of each group is detected, and the results are shown ...

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Abstract

The invention belongs to the field of microorganisms, and discloses an artificially induced Geobacillus thermoglucosidase and its application in cellulose degradation. The applicant has isolated a cellulase enzyme from pig carcasses and samples of straw composting at high temperature. The most active strain Geobacillus thermodenitrificans , after high temperature acclimation and ultraviolet mutagenesis, a strain of Geobacillus thermoglucosidase with high cellulase production was obtained. Geobacillus glucosidase BGSC95A1. A variety of carbon sources can be used, and it can degrade a variety of celluloses, and the concentration of cellulase enzymes in the fermentation broth produced by fermentation is high.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to an artificially induced Geobacillus thermoglucosidase and its application in cellulose degradation. Background technique [0002] As the most abundant and least utilized resource in the biosphere, cellulose can be hydrolyzed by microorganisms with the help of a multi-enzyme system (Hahn- al 2006). Cellulase is a complex enzyme with synergistic effect, mainly including endoglucanase (EC3.2.1.4endo-1 4-β-D-glucanase), exoglucanase (EC3.2.1.91, exo -1 4-β-D-glucanase) and β-glucosidase (EC.3.2.1.21, β-glucosidase). Three enzymes coordinately participate in the degradation of natural cellulose (Li Na 2006), endoglucanase mainly works in the non-crystalline region of cellulose molecules, hydrolyzes β-1,4 glycosidic bonds, and makes long-chain cellulose molecules. The chains become small molecules of cellulose, creating more cleavage sites for exoglucanases and beta-glucos...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/42C12P19/02C12P19/14C12P19/12C12R1/01
CPCC12N9/2437C12P19/14C12P19/12C12P19/02C12R2001/01C12N1/205Y02W30/40
Inventor 郑嵘韩珍珍杨旭晨刘建华李宇飞蒋思文柴进
Owner HUAZHONG AGRI UNIV
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