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High-yield glutamine transaminase strain and application thereof

A technology of transaminase strains and glutamine, applied in the direction of bacteria, microorganisms, biochemical equipment and methods, etc., can solve the problems of incomplete activity and affect the application of glutamine transaminase, so as to reduce production costs and improve production efficiency Effect

Active Publication Date: 2019-10-22
TAIXING DONGSHENG FOOD TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The activities of MTGase from different Streptomyces genus and even the MTGase secreted by Streptomyces of the same genus and different species are not exactly the same, and there are great differences, which affect the application of transglutaminase in the later stage

Method used

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  • High-yield glutamine transaminase strain and application thereof
  • High-yield glutamine transaminase strain and application thereof
  • High-yield glutamine transaminase strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (1) Gao's No. 1 culture medium for preparing the high-yield transglutaminase bacterial strain Streptomyces mobaraensis of the present invention, comprises the raw material composition of following weight ratio: agar: 2%; Starch: 2%; Potassium nitrate: 1%; Potassium: 0.5%; Sodium Chloride: 0.5%; Magnesium Sulfate Heptahydrate: 0.5%; Ferrous Sulfate Heptahydrate: 0.01%; pH: 7.4.

[0032] The method for preparing the above-mentioned Gao's No. 1 medium may further comprise the steps:

[0033] ①. Weigh 2% agar, 2% starch, 1% potassium nitrate, 0.5% dipotassium hydrogen phosphate, 0.5% sodium chloride, 0.5% magnesium sulfate heptahydrate, and 0.01% ferrous sulfate heptahydrate.

[0034] ②. After making up the volume with water, adjust the pH to 7.4, and sterilize under high temperature and high pressure at 121°C for 20 minutes.

[0035] ③. Pour the prepared culture medium into each petri dish at a volume of 20mL / dish (the diameter of the petri dish is 90mm) and wait to cool ...

Embodiment 2

[0038] (2) prepare the seed culture medium and fermentation medium of the high-yield transglutaminase bacterial strain Streptomyces mobaraensis of the present invention, comprise the raw material composition of following weight ratio: glycerol 2%; Yeast extract 0.6%; Fish meal peptone 2.5%; Magnesium sulfate heptahydrate 2%, dipotassium hydrogen phosphate 2%, and the rest is pure water; pH is 7.4.

[0039] The method for preparing above-mentioned seed medium, fermentation medium:

[0040] ①. Weigh 2% agar, 2% glycerin, 0.6% yeast extract, 2.5% fish powder peptone, 2% dipotassium hydrogen phosphate, 2% magnesium sulfate heptahydrate; among them, add 1% accelerator to each bottle of fermentation medium.

[0041] ②. After making up the volume with water, adjust the pH to 7.4, and sterilize under high temperature and high pressure at 121°C for 20 minutes.

[0042] ③. Cool the prepared medium to room temperature for later use.

Embodiment 3

[0043] Embodiment 3 prepares the reagent of high-yield transglutaminase bacterial strain of the present invention:

[0044] (1) Preparation of reagents A and B

[0045] Reagent A (reaction solution): containing 0.2M Tris-HCl, 0.1M hydroxylamine hydrochloride, 0.01M reduced glutathione, 0.03M Na-CBZ-Gln-Gly, pH 6.0.

[0046]Reagent B (Stop Solution): Mix the following three components in a volume ratio of 1:1:1. 3mol / L HCl, 12% trichloroacetic acid (W / V), 5% ferric chloride (W / V).

[0047] (2) Carry out enzyme activity detection to bacterial strain of the present invention by standard hydroxamate assay method, and its step is: get 2ml reagent A liquid and reagent B liquid respectively and add in ① and ② two grinding mouth test tubes respectively, 37 ℃ of insulations 15 Minutes, add 0.2ml enzyme solution, react for 10 minutes, add 2ml reagent B solution to ① test tube to stop the reaction, add 2ml reagent A solution to ② reagent test tube, use ② test tube reaction solution as ...

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Abstract

The invention relates to the technical field of microorganism mutation breeding, in particular to a high-yield glutamine transaminase strain. The high-yield glutamine transaminase strain (YRZ170817NTG31) is preserved in the China Center For Type Culture Collection and has the preservation number of 15353 and the preservation date of February 12th, 2018. The high-yield glutamine transaminase straincan has the maximum enzyme activity after 24 hours of seed culturing and 48-56 hours of fermentation culturing and has the enzyme activity of 20-40 U / mL. The strain has the advantage of being high inenzyme activity.

Description

technical field [0001] The invention relates to the technical field of microbial mutation breeding, in particular to a mutagenesis method and a screening method for high-throughput screening of high-yield transglutaminase strains, high-yield transglutaminase strains and applications thereof. Background technique [0002] Transglutaminase (TGase or TG for short) is a monomeric protein acyltransferase with an active center and a molecular weight of about 38,000 composed of 331 amino groups. Transglutaminase can catalyze the cross-linking between proteins and produce the following effects: modify the protein, improve the plasticity, water holding capacity, water solubility and functionality of the modified protein; protect the lysine in food Acid is protected from various chemical reactions; it can be used to embed lipids or fat-soluble substances; it can form heat-resistant and water-resistant films; it does not require heat treatment during the formation of gels; Cross-linki...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/01C12R1/465
CPCC12N1/20C12N15/01C12R2001/465C12N1/205
Inventor 步国建张玉如甘李寇雪蕊和超楠刘艳贺晓洁
Owner TAIXING DONGSHENG FOOD TECH
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