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Culture medium for screening high-yield glutamine transaminase strain in high-throughput mode

A technology of glutamine and transaminase bacteria, which is applied in the direction of acyltransferase, transferase, enzyme, etc., can solve the problems of unsatisfactory accuracy and efficiency, and achieve the effects of shortening the overall cycle, reducing costs and simplifying the process

Active Publication Date: 2016-08-03
EAST CHINA NORMAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The screening of traditional high-yielding transglutaminase strains relies on measuring the enzyme activity after fermentation in a 96-well plate, but its accuracy and efficiency are not ideal

Method used

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  • Culture medium for screening high-yield glutamine transaminase strain in high-throughput mode
  • Culture medium for screening high-yield glutamine transaminase strain in high-throughput mode

Examples

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Embodiment 1

[0027] Embodiment 1 prepares the medium of novel high-throughput screening high-yield transglutaminase strain of the present invention, comprises the raw material composition of following weight ratio:

[0028] Enzymatic Hydrolyzed Casein: 1%

[0029] Agar: 2%

[0030] Starch: 2%

[0031] Potassium nitrate: 1%

[0032] Dipotassium hydrogen phosphate: 0.5%

[0033] Sodium chloride: 0.5%

[0034] Magnesium sulfate heptahydrate: 0.5%

[0035] Ferrous sulfate heptahydrate: 0.01%

[0036] Cresol red: 0.001%

[0037] pH: 7.4

[0038] The method for making the culture medium for novel high-throughput screening of high-yielding transglutaminase strains as described above:

[0039] 1. Weigh 1% enzymatically hydrolyzed casein, 2% agar, 2% starch, 1% potassium nitrate, 0.5% dipotassium hydrogen phosphate, 0.5% sodium chloride, 0.5% magnesium sulfate heptahydrate, 0.01% sulfurous acid heptahydrate iron.

[0040] 2. After making up the volume with water, adjust the pH to 7.4. Aft...

Embodiment 2

[0045] Embodiment 2 prepares the medium of novel high-throughput screening high-yielding transglutaminase strain of the present invention, comprises the raw material composition of following weight ratio:

[0046] Gelatin: 2%

[0047] Agar: 2%

[0048] Starch: 2%

[0049] Potassium nitrate: 1%

[0050] Dipotassium hydrogen phosphate: 0.5%

[0051] Sodium chloride: 0.5%

[0052] Magnesium sulfate heptahydrate: 0.5%

[0053] Ferrous sulfate heptahydrate: 0.01%

[0054] Phenol red: 0.01%

[0055] pH: 7.4

[0056] The method for making a novel high-throughput screening medium for high-yielding transglutaminase strains as described above:

[0057] 1. Weigh 2% gelatin, 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.

[0058] 2. After making up the volume with water, adjust the pH to 7.4, sterilize under high temperature and high pressure at 121°C...

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Abstract

The invention relates to the field of microbial breeding and screening and discloses a novel culture medium for screening high-yield glutamine transaminase strains in a high-throughput mode. The culture medium comprises, by weight, 0.1-2% of glutamine transaminase substrates, 1.5-4% of agar, 2% of starch, 1% of nitrate of potash, 0.5% of dipotassium phosphate, 0.5% of sodium chloride, 0.5% of magnesium sulfate heptahydrate, 0.01% of iron sulfate heptahydrate, 0.0001-0.01% of an acid-base indicator. The pH ranges from 6 to 8. The culture medium has the advantages of being short in cycle, capable of screening a great number of bacterial strains at the same time and free of any mark so that the high-yield glutamine transaminase strains can be obtained through direct screening when used for screening the high-yield glutamine transaminase strains. The invention further discloses a use method of the culture medium, and the bacterial strains to be selected are cultured for 3-5 days on the culture medium and used for screening high-yield bacterial strains.

Description

technical field [0001] The invention relates to the technical field of microbial breeding, in particular to a novel high-throughput screening medium for high-yield transglutaminase strains and a preparation method thereof. Background technique [0002] Transglutaminase is a biocatalyst with a wide range of uses, and its commercial value has also been fully demonstrated in the food industry. Obtaining high-yielding transglutaminase strains is a key point in breeding. It can be said that a high-throughput screening medium has great advantages in obtaining high-yielding strains. Traditional screening of high-yielding transglutaminase strains relies on measuring enzyme activity after fermentation in a 96-well plate, but its accuracy and efficiency are not ideal. Contents of the invention [0003] The technical problem to be solved by the present invention is to provide a short-period culture medium capable of simultaneously screening a large number of bacterial strains, as we...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/02C12N1/20C12N9/10C12R1/465
CPCC12N1/02C12N1/20C12N9/1044C12Y203/02013
Inventor 金明飞常忠义高红亮黄劲舸宋佳雯步国建
Owner EAST CHINA NORMAL UNIV
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