L-d-glutamic oxidase with substrate specificity and alpha-oxoglutarate produced by catalysis of same

A technology of glutamic acid oxidase and sodium glutamate, which is applied in the direction of enzymes, microorganisms, and methods based on microorganisms, can solve problems such as environmental pollution and raw material sources, and achieve reduced operation difficulty, low production costs, and simple production processes Effect

Inactive Publication Date: 2013-03-27
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the industrial production adopts the organic synthesis method, which involves a series of complex chemical reaction processes, which causes a series of problems such as raw material sources and environmental pollution.
At present, there is no report on the production of α-KG by biocatalysis at home and abroad.

Method used

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  • L-d-glutamic oxidase with substrate specificity and alpha-oxoglutarate produced by catalysis of same
  • L-d-glutamic oxidase with substrate specificity and alpha-oxoglutarate produced by catalysis of same
  • L-d-glutamic oxidase with substrate specificity and alpha-oxoglutarate produced by catalysis of same

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0025] 1. L-glutamate oxidase substrate specificity experiment:

[0026] According to the Trinder reaction, the L-type amino acid solution with a concentration of 11 mg / mL was prepared with pH 7.0 phosphate buffer solution for color reaction measurement. After an accurate reaction at 30°C for 30 minutes, it was boiled at 100°C for 5 minutes and measured at 550 nm.

[0027]

[0028] 2. The effect of different pH on the catalytic transformation of L-glutamic acid oxidase:

[0029] Prepare the fermentation broth containing L-glutamic acid oxidase in different pH range gradients from 5.5 to 10.5, add 20g / L sodium L-glutamate solution, transform for 12 hours, and measure the content of α-ketoglutaric acid in the fermentation broth . As a preferred version of the present invention, 8.5 is selected as the conversion pH, at which time the conversion rate is the highest and is closest to the pH of the fermentation broth.

[0030]

[0031]

[0032] 3. Put 40mL of enzyme-conta...

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Abstract

The invention discloses L-d-glutamic oxidase with substrate specificity and alpha-oxoglutarate produced by catalysis of the same, and belongs to the field of enzyme-method catalytic production of fine chemicals. The invention has the advantages that the L-d-glutamic oxidase which only has the substrate specificity to L-glutamic acid, L-sodium glutamate and glutamine is utilized, 20g/L L-sodium glutamate solution is added in fermented liquid which is fermented for 60 hours and contains the L-d-glutamic oxidase, carrying out conversion for 12 hours under the ventilating condition and the conditions that the temperature is 30 DEG C, the pH is 8.5 and the 5% (v/v) isopropyl alcohol, and measured by a high-performance liquid chromatography, the content of alpha-oxoglutarate reaches 14.5g/L.

Description

technical field [0001] The invention relates to a production method for converting L-glutamic acid into α-ketoglutaric acid in one step by microbial enzymatic method, and belongs to the field of fine chemicals produced by catalytic method. Background technique [0002] α-ketoglutarate (α-KG), also known as α-glutaric acid, 2-oxoglutarate or α-oxoglutarate, the molecular formula is C 5 h 6 o 5 , the relative molecular weight is 146.1, α-KG is white or off-white crystal or crystalline powder, colorless, easily soluble in water, plays an important role in the metabolism of microbial cells, and is an important part of the tricarboxylic acid cycle (TCA). One of the intermediate products, it participates in the synthesis of amino acids, proteins, vitamins and energy metabolism in organisms. α-KG has important application prospects in various fields. As a nutritional enhancer, it is widely used in industries such as food, medicine, organic synthesis, cosmetics and feed. [0003...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/06C12P7/50C12R1/465
Inventor 刘立明牛盼清林小宝
Owner JIANGNAN UNIV
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