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Method and special-purposed strain used for producing alpha-ketoglutaric acid

A technology of ketoglutaric acid and pyruvic acid is applied in the field of producing α-ketoglutaric acid, which can solve the problems such as α-ketoglutaric acid of Absidia agaricus that have not yet been seen, and achieve good scientific research value and industrial application prospect. , the optimization of fermentation conditions, the effect of a wide range of substrate sources

Active Publication Date: 2014-05-07
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are few reports on the production of α-ketoglutarate by fermentation of fungi other than yeast at home and abroad, and there is no research on the production of α-ketoglutarate by Lichtheimia corymbifera

Method used

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  • Method and special-purposed strain used for producing alpha-ketoglutaric acid
  • Method and special-purposed strain used for producing alpha-ketoglutaric acid
  • Method and special-purposed strain used for producing alpha-ketoglutaric acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1, the isolation and identification of Absidia coliformis D1

[0032] 1. Isolation of Absidia codica D1

[0033] Absidia codica D1 was screened from distiller's yeast samples.

[0034] Primary screening medium: add 1.0g / L sodium deoxycholate and 0.1g / L bromocresol green to PDA medium, natural pH.

[0035] Incline medium (PDA medium): potato 200g / L, glucose 20g / L, agar 20g / L, natural pH.

[0036] Fermentation medium: glucose 80g / L, (NH 4 ) 2 SO 4 1g / L, KH 2 PO 4 1.36g / L, MgSO 4 ·7H 2 O 0.2g / L, FeSO 4 ·7H 2 O 0.01g / L, NaCl 2g / L, corn steep liquor 0.5g / L, CaCO 3 30g / L (sterilized separately), natural pH.

[0037] The distiller's yeast samples were diluted and spread on the separation medium, and cultured in a constant temperature incubator at 37°C. When colonies grow out of the separation medium, inoculate the strains into the primary screening medium, inoculate all the strains that can turn the indicator medium yellow around the colonies into th...

Embodiment 2

[0043] Example 2. Optimization of conditions for the production of α-ketoglutarate by shake flask fermentation using sucrose as a substrate

[0044] 1. Screening of carbon source in fermentation medium

[0045] 1. Screening of carbon sources

[0046] The fermentation medium is obtained by distilling the following substances to 1L (natural pH): carbon source 60g, (NH 4 ) 2 SO 4 1g, KH 2 PO 4 1.36g, MgSO 4 ·7H 2 O 0.2g, FeSO 4 ·7H 2 O 0.01g, NaCl 2g, corn steep liquor 0.5g, CaCO 3 30g. The following seven carbon sources were used: glucose, sucrose, maltose, raffinose, glutinous rice starch, corn starch, and soluble starch.

[0047] Experimental method: in 250mL Erlenmeyer flask, pack 50mL fermentation medium, inoculate 200 μ L of the spore liquid of Absidia codica D1 (the concentration of spores in the spore liquid is 1×10 8 spores / mL), cultured on a reciprocating shaker in a constant temperature air bath at 37°C at 200r / min for 72h; take the fermentation broth, c...

Embodiment 3

[0100] Example 3. Optimization of conditions for the production of α-ketoglutarate by shake flask fermentation using pyruvate as a substrate

[0101] 1. Screening of the amount of pyruvate added in the fermentation medium

[0102] The fermentation medium is obtained by distilling the following substances to 1L (natural pH): pyruvate, (NH 4 ) 2 SO 4 1g, KH 2 PO 4 1.36g, MgSO 4 ·7H 2 O 0.2g, FeSO 4 ·7H 2 O 0.01g, NaCl 2g, corn steep liquor 0.5g, CaCO 3 60g. The following 8 adding amounts of pyruvic acid were adopted respectively: 10g, 20g, 30g, 40g, 50g, 60g, 70g, 80g.

[0103] Experimental method: in 250mL Erlenmeyer flask, pack 50mL fermentation medium, inoculate 200 μ L of the spore liquid of Absidia codica D1 (the concentration of spores in the spore liquid is 1×10 8 spores / mL), cultured on a reciprocating shaker in a constant temperature air bath at 37°C at 200r / min for 72h; take the fermentation broth, centrifuge at 12000r / min for 10min, absorb the supernatan...

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Abstract

The invention provides a method and a special-purposed strain used for producing alpha-ketoglutaric acid. Lichtheimia corymbifera D1 provided by the invention has a collection number of CGMCC NO.4974. The lichtheimia corymbifera D1 provided by the invention has good strain stability, good alpha-ketoglutaric acid production capacity, relatively low fermentation cost (substrates have wide source and low cost), and good industrialized application prospect. The invention also optimizes the fermentation conditions of lichtheimia corymbifera D1. The method provided by the invention is used for producing alpha-ketoglutaric acid, and has good scientific research value and industrial application prospect.

Description

technical field [0001] The invention relates to a method for producing α-ketoglutarate and a special strain thereof, in particular to a method for producing α-ketoglutarate by using sucrose and pyruvic acid. Background technique [0002] α-ketoglutarate (α-KG), also known as α-gluconic acid, the molecular formula is C 5 h 6 o 5 . α-ketoglutaric acid is white or off-white crystalline powder, easily soluble in water. α-ketoglutaric acid mainly exists in various types of cheese, beer and coffee, and is a natural equivalent spice. α-ketoglutarate plays an important role in the metabolism of microbial cells and is an important precursor for the synthesis of amino acids, proteins, and vitamins. α-ketoglutaric acid is widely used as a nutritional enhancer in food, medicine, organic synthesis, cosmetics and feed industries. [0003] With the continuous expansion of the scope of application and the continuous growth of market demand, the industrial production of α-ketoglutaric ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P7/50C12R1/645
Inventor 闫巧娟徐忠义江正强蔡威
Owner CHINA AGRI UNIV
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