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Two-stage pH (Potential Of Hydrogen) control method for high output of alpha-ketoglutaric acid

A ketoglutaric acid, high-yield technology, applied in the field of two-stage pH control strategy, can solve the problem that the production intensity cannot reach industrialization

Active Publication Date: 2012-07-18
JIANGNAN UNIV +1
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Problems solved by technology

Our research team screened a strain of Y.lipolytica CCTCC NO.M207143 with high α-ketoglutaric acid production from the soil. After a series of single-factor optimization, it was found that the production intensity of this strain still cannot meet the needs of industrialization.

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0024] Example 1 Two-stage pH control strategy for excessive accumulation of α-ketoglutarate

[0025] Wash the spores of Yarrowia lipolytica cultured at 28°C for 10 days on the spore-forming medium with sterile water, transfer the spore suspension to the previous culture medium at a volume ratio of 4%, and cultivate at 28°C at 400rpm for 40h; at 10% The volume ratio of the seed to the fermentation medium before transfer, 28 ℃, 400rpm to maintain the ventilation rate of 1.5vvm, add 20g / L CaCO 3 Use the pH buffer of the fermentation broth to cultivate until the pH of the fermentation broth drops to 2.5, and then use 4mol / L NaOH solution to maintain the pH of the fermentation broth at 2.5 and cultivate for 168h.

[0026] The two-stage pH control strategy is compared with the matched group: (1) by-product pyruvate output is 36.9g / L in the comparative example 1 (maintaining pH4.5), and the comparative example 2 (CaCO 3 As buffering agent), by-product pyruvic acid output is 31.8g / L...

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Abstract

The invention discloses a two-stage pH (Potential Of Hydrogen) control method for the high output of alpha-ketoglutaric acid, which belongs to the field of fermentation engineering. The method adopts two stages of pH control to control the process of fermentation, and adopts Yarrowia lipolytica CCTCC NO.M207143 as a producing strain, CaCO3 is adopted as a pH buffer in the early stage of fermentation, NaOH is used for keeping the pH at 2.5 in the later stage, a great deal of Alpha-ketoglutaric acid is accumulated, and the output of alpha-ketoglutaric acid on the laboratory scale reaches 53.4g / L; in comparison with constant pH equal to 2.5, the output of alpha-ketoglutaric acid is increased by 142.7 percent; in comparison with only CaCO3 as the pH buffer, the output of alpha-ketoglutaric acid is increased by 32.5 percent; in the later stage of fermentation, the low pH value can decrease the probability of contamination, the dosage of NaOH is reduced, and thereby the extraction of downstream product is benefited; and the method has a broad industrial application prospect.

Description

technical field [0001] The present invention relates to a high-yield α-ketoglutaric acid fermentation strategy, especially a two-stage pH control strategy. Background technique [0002] α-ketoglutaric acid (α-KG) is an important dicarboxylic acid widely used in food, medicine, fine chemical and other industrial production. At present, the production method of α-ketoglutaric acid is mainly chemical synthesis. The chemical synthesis method has too many by-products, and the toxic substance cyanide and residual organic compounds in the by-products increase the difficulty and cost of extraction of α-ketoglutarate, and limit the application range of α-ketoglutarate. Microbial fermentation has the advantages of high purity and environmental friendliness, but the fact that the production intensity of α-ketoglutarate produced by fermentation is low makes industrial production choose chemical synthesis without any suspense. [0003] Previously, scholars at home and abroad studied th...

Claims

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Application Information

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IPC IPC(8): C12P7/50C12R1/645
Inventor 陈坚周景文余宗钟堵国成
Owner JIANGNAN UNIV
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