Glutamic acid decarboxylase recombinant bacterium, and construction method and application thereof

A technology of glutamic acid decarboxylase and construction method, which is applied in the directions of application, recombinant DNA technology, botanical equipment and methods, etc., can solve the problem of low L-glutamic acid decarboxylase enzyme activity, difficulty in downstream separation and purification, and complicated extraction process and other problems, to achieve the effect of short reaction time, wide source of raw materials and high enzyme activity
CN103484419AActive Publication Date: 2014-01-01NANJING UNIV OF TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
NANJING UNIV OF TECH
Publication Date
2014-01-01

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Abstract

The invention discloses a glutamic acid decarboxylase recombinant bacterium, which is an Escherichia coli introduced with L-glutamic acid decarboxylase GAD; and the L-glutamic acid decarboxylase GAD gene is from yeast Kodamaea ohmeri NH-9 and has a nucleotide sequence shown as a SEQ ID No:1. The invention also discloses the construction method and application of the above glutamic acid decarboxylase recombinant bacterium. The invention has the following advantages: (1) hydrochloric acid pyridoxal has a price half of that of 5 ' -phosphoric acid pyridoxal, so as to reduce technology cost; (2) the method has high enzyme activity, short reaction time, mild conditions, substrate concentration up to 510 g / L and concentration of a product gamma-aminobutyric acid up to 352 g / L; (3) the bacteria and the buffer after rotary evaporation can be reused, and the conversion rate is 100%, and the yield is no less than 99%; and (4) the technology is simple, green and can realize a product purity higher than 99%.
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Description

technical field

[0001] The invention relates to a glutamic acid decarboxylase recombinant bacterium and its construction method and application, belonging to the technical field of genetic engineering. Background technique

[0002] γ-GABA is the product of the decarboxylation of L-glutamic acid ( figure 1 ), its chemical name is γ-aminobutyric acid (abbreviated as γ-GABA), and its molecular formula is NH 2 CH 2 CH 2 CH 2 COOH is very soluble in water, and can exist in the form of variable molecular structure in the solution, which can be stretched into a line or form a ring like proline. γ-GABA is an amphiphilic molecule with an isoelectric point of 7.3 close to the physiological pH value. The finished product is colorless to white odorless needle crystal or crystalline powder.

[0003] γ-GABA has a wide range of uses and can be used in food, feed, medicine and chemical industries. It has the following physiological functions: 1 soothe the nerves and antidepressant; 2...

Claims

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