Method for microbial fermentation synthesis of ª‡-ketoglutaric acid

A technology for microbial fermentation, ketoglutaric acid, used in microorganism-based methods, biochemical equipment and methods, microorganisms, etc.

Inactive Publication Date: 2006-06-14
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, microbial fermentation and synthesis of α-ketoglutarate has not been reported in the literature.

Method used

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  • Method for microbial fermentation synthesis of ª‡-ketoglutaric acid
  • Method for microbial fermentation synthesis of ª‡-ketoglutaric acid
  • Method for microbial fermentation synthesis of ª‡-ketoglutaric acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Sterilize the medium in a 7L fermenter, inoculate microbial seeds according to 10% (volume ratio), after inoculation and mixing, transfer 50mL of inoculated medium aseptically to a sterilized 500mL Erlenmeyer flask, and add 40g / L calcium carbonate was cultured in shake flasks, cultured at 30°C and 200rpm for 48h, and the concentration of α-ketoglutaric acid was measured to be 15.8g / L, C PYR / C KG is 3.1.

Embodiment 2

[0051] The inoculation and culture conditions were the same as in Example 1, and 20-40 μg / L of biotin was added. It was found that the concentration of α-ketoglutarate increased with the increase of biotin concentration, and C PYR / C KG slowing shrieking. When the total concentration of biotin in the medium reaches 60 μg / L, the concentration of α-ketoglutarate is 23.5 g / L, C PYR / C KG is 1.47.

Embodiment 3

[0053] Put 4L of fermentation medium in a 7L fermenter and then sterilize it, add Torulopsis glabrata according to 10% (volume ratio), add 40g / L calcium carbonate and 60μg / L biotin in the medium. After culturing at 30°C, ventilation rate 3L / (L·min), rotation speed 400rpm for 60h, the concentration of α-ketoglutaric acid was 38.4g / L, C PYR / C KG is 0.72.

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Abstract

The invention discloses a method for microbial fermentative synthesis of α-ketoglutaric acid, in which calcium carbonate is added to a culture medium and the concentration of biotin is increased to promote the generation of a large amount of α-ketoglutarate during pyruvate fermentation. During the fermentative production of pyruvate by Torulopsis glabrata CCTCC M202019, delaying the addition of calcium carbonate inhibited the formation of α-KG and increased the carbon molar ratio of pyruvate to α-KG (C PYR / C KG ), and increasing the concentration of calcium carbonate in the medium will promote the accumulation of a large amount of α-KG, and when the concentration of calcium carbonate is 40g / L, it is most conducive to the formation of α-KG. Keep the concentration of calcium carbonate in the medium constant, increase the concentration of biotin in the medium, promote the concentration of α-KG to rise continuously and C PYR / C KG The value decreased continuously, and the accumulation of α-KG was 23.5g / L when the biotin concentration was 60μg / L. When there is Ca 2+ When present, the activity of intracellular pyruvate carboxylase can be increased by 40%, while the activity of pyruvate dehydrogenase system does not change significantly. Ca in medium 2+ The increase of the concentration of biotin and biotin can significantly increase the activity of pyruvate carboxylase, so that T. glabrata shifts from fermentative production of pyruvate to synthesis of high-concentration α-KG.

Description

technical field [0001] The invention relates to a method for synthesizing α-ketoglutaric acid by microbial fermentation. Background technique [0002] α-ketoglutaric acid (α-ketoglutaric acid, referred to as α-KG), chemical name: 2-oxo-1,5 glutaric acid, structural formula is [0003] [0004] It is a white or off-white powder substance, easily soluble in water, and is widely used as an organic intermediate in the production of organic synthesis; it is an important biochemical reagent, and it can also be used as a supporting reagent for the determination of liver function. Nutritional fortifiers are widely used in: improving the body's immunity; reducing the body loss of postoperative patients and long-term patients; serving as precursors for the synthesis of aminobutyric acid and glutamic acid in the brain; related to energy production in the body. As a component of sports nutrition drinks, it can promote aerobic energy supply; improve athletes' insulin levels, promote ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/50C12R1/85
Inventor 陈坚李寅刘立明伦世仪
Owner JIANGNAN UNIV
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