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Bacillus amyloliquefaciens and application thereof to fermentation production of glutaminase

A technology for producing glutamine and glutaminase, applied in the directions of hydrolase, microorganism-based methods, enzymes, etc., can solve the problems of low yield, limited fermentation mode, low fermentation enzyme activity, etc., and achieves easy extraction and easy extraction. Product industrial production, high purity effect

Active Publication Date: 2021-02-02
安琪酶制剂(宜昌)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] For this reason, the technical problem solved by the present invention is: the production of glutaminase mainly adopts solid-state fermentation production, the activity of fermented enzyme is low, extraction is relatively difficult, and it is difficult to be fermented. Way limited, output is not high

Method used

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  • Bacillus amyloliquefaciens and application thereof to fermentation production of glutaminase
  • Bacillus amyloliquefaciens and application thereof to fermentation production of glutaminase
  • Bacillus amyloliquefaciens and application thereof to fermentation production of glutaminase

Examples

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Embodiment 1

[0055] The acquisition of embodiment 1 bacterial strain

[0056] (1) Isolation strain: the strain is isolated from the soil around the factory producing fermented bean curd or soy sauce. The specific operation steps are:

[0057] Sample collection: collect samples from the soil around the fermented bean curd or soy sauce factory or from the semi-finished fermented bean curd or fermented soy sauce;

[0058] Microbial enrichment: place the collected samples in a constant temperature environment at 37°C for more than 48 hours until dry; take an appropriate amount of dry samples and inoculate them into fresh medium for enrichment culture; the medium formula is glucose 1%, yeast powder 2 %, glutamine 0.06%, dipotassium hydrogen phosphate 0.05%, potassium dihydrogen phosphate 0.2% and magnesium chloride 0.1%, the balance is water

[0059] The enriched samples were plate cultured after serial dilution to obtain single colonies;

[0060] (2) Mutagen strains

[0061] Adopt ARTP muta...

Embodiment 2-1

[0078] Embodiment 2-1 fermentation produces glutaminase

[0079] (1) Strain activation: select glycerol tube strains and activate them in a fresh plate culture medium at 37°C for 18 hours to obtain a single colony. The components of the plate culture medium are: based on 100L of water, yeast powder 5g / L, peptone 10g / L, sodium chloride 10g / L;

[0080] (2) Seed preparation: select 1-2 single bacterium colonies after activation and transfer in the Erlenmeyer flask with fresh seed culture medium, (its composition and content are the same as the culture medium of step (1)) (500mL Fill 100mL feed solution in the Erlenmeyer flask), prepare seed solution, culture condition: 37 ℃, 200rpm culture 10h;

[0081] (3) Seed tank cultivation: the inoculum size is 0.3% by volume to inoculate the seed solution obtained in step (2) into 5L seed tanks containing the seed medium for cultivation, the cultivation temperature is 37°C, the rotating speed is 100rpm, and the ventilation ratio 1:0.8, ...

Embodiment 2-2

[0118] Embodiment 2-2 fermentation produces glutaminase

[0119] (1) Strain activation: select the glycerol tube strain and activate it in a fresh plate medium at 37°C for 18 hours to obtain a single colony. The components of the plate medium are the same as the components and contents of (1) in Example 2-1 same;

[0120] (2) Seed preparation: 1-2 single bacterium colonies activated in the selection step (1) are transferred to fresh seed medium (which has the same composition and content as the embodiment 2-1 step (2)) In the Erlenmeyer flask (250mL Erlenmeyer flask is equipped with 50mL feed liquid), prepare seed liquid, culture condition: 37 ℃, 200rpm culture 12h;

[0121] (3) Seed tank cultivation: the inoculum size is 0.2% by volume to inoculate the seed solution obtained in step (2) into a 5L seed tank containing the seed medium for cultivation, the cultivation temperature is 37°C, the rotating speed is 200rpm, the ventilation 1:1.1, cultured for 10 hours, wherein, the ...

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Abstract

The invention relates to the field of microorganism screening and fermentation, in particular to bacillus amyloliquefaciens and application thereof to fermentation production of glutaminase. The invention provides a strain for producing glutaminase, the strain is Bacillus amyloliquefaciens ESP827 which is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC No:M2018828. The invention further provides a method using the strain to perform liquid state fermentation for producing glutaminase. The fermentation enzyme activity of the glutaminase obtained bythe method is far higher than that of currently reported glutaminase, the fermentation enzyme activity is 120-158U / mL, the liquid state fermentation mode is easier for industrial production of products, the fermentation enzyme activity purity is higher, and downstream extraction is facilitated.

Description

technical field [0001] The invention relates to the field of microbial screening and fermentation, in particular to a bacillus amyloliquefaciens and its application in fermentation production of glutaminase. Background technique [0002] Glutaminase is a kind of amidase, which can specifically catalyze the hydrolysis of L-β-glutamine to generate L-glutamic acid and ammonia. Glutamic acid is an amino acid with umami taste, and sodium glutamate is recognized as a safe and healthy seasoning. Adding glutaminase to various products such as yeast extract, soy sauce, vegetable protein hydrolyzate, meat and fish protein hydrolyzate, miso soup, fruit and vegetable hydrolyzate can increase the content of glutamic acid, improve and improve the composition of amino acids points, increase and strengthen the taste performance of the product. In addition, glutaminase can also inhibit the conversion of glutamine in protein raw materials to tasteless pyroglutamic acid, and improve the util...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/80C12R1/07
CPCC12N9/80C12N1/20C12Y305/01002C12R2001/07C12N1/205
Inventor 喻晨俞学锋李知洪李汉文胡悦栾春艳姚鹃余华顺
Owner 安琪酶制剂(宜昌)有限公司
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