Method for implementing excessive accumulation of alpha-oxoglutarate by regulation and control of carbon metabolism flow with assist factor

A technology of ketoglutarate and cofactor, applied in the field of excessive accumulation of α-ketoglutarate

Inactive Publication Date: 2008-04-02
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on how vitamins and metal ions affect carbon metabolism and the flow direction distribution of carbon metabolism in industrial microorganisms

Method used

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  • Method for implementing excessive accumulation of alpha-oxoglutarate by regulation and control of carbon metabolism flow with assist factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 improves vitamin B 1 Concentration increases pyruvate dehydrogenase (PDH) pathway metabolic flux

[0034] Vitamin B 1 When the concentration increased to 0.04mg / L, the production of α-KG reached a maximum of 10.3g / L, the production of pyruvate decreased to 58.6g / L, and the carbon molar ratio of pyruvate to α-KG (CPYR / CKG) was 3.9. Continue to increase the vitamin B in the medium 1 When the concentration is high, the production of α-KG will gradually decrease.

Embodiment 2

[0035] Example 2 Increasing the Biotin Bio Concentration Increases the Metabolic Flux of the Pyruvate Carboxylase (PC) Pathway

[0036] When the biotin Bio concentration in the medium increased to 0.06mg / L, the α-KG production reached the maximum value of 14g / L, the pyruvate production dropped to 52.6g / L, and the carbon molar ratio of pyruvate to α-KG (CPYR / CKG) was 3.74.

Embodiment 3

[0037] Example 3 Calcium Carbonate Effects on Carbon Metabolic Flow Direction and Flux Size

[0038] As shown in the table below, when the pH was not adjusted in shake flasks and fermenters, cell growth was weak and acid production was minimal. In shake flasks with CaCO 3 When used as a buffer, a higher concentration (6.8 g / L) of α-KG was produced. In fermentor culture, if the pH is adjusted with NaOH, the concentration of α-KG in the fermentation broth is very low. But if instead CaCO 3 When the pH was adjusted, the concentration of α-KG in the fermented liquid increased by 8 times compared with that when the pH was adjusted with NaOH. When the calcium carbonate concentration in the culture system was 60g / L, the accumulation of α-KG reached the maximum value (12.8g / L).

[0039] Fermentation controls in shake flasks and fermenters

[0040] cell

[0041] Description: C PYR / C KG It is the ratio of the number of carbon moles of pyruvate to the number of carbon...

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Abstract

A method with adjuvant factor to regulate the carbon metabolism to accomplish the overdose accumulation of alpha-ketoglutarate (alpha-KG) is provided, which belongs to the technical field of fermentation process optimized by the strategy of adjuvant factor regulation. The invention adopts the method of adjuvant factors including vitamins and metal ions, etc. which are added into the fermentation process with the sub-proper amount, and then the pathway flux of PDH and PC are increased selectively to accomplish the overdose accumulation of alpha-KG. During the process of the production of alpha-KG through fermentation of T.glabrata, the concentration of vitamin B1 is raised to increase the accumulated alpha-KG to 10.3g / L; then the biotin concentration is raised to increase the accumulated alpha-KG to 43.7g / L, and the concentration of pyruvic acid is decreased to 21.8g / L. The strategy also has the significance to guide the production of the other fermented products.

Description

technical field [0001] The invention relates to a method for cofactor regulating carbon metabolic flow to realize excessive accumulation of α-ketoglutarate (α-KG). By studying the effect of cofactors (vitamins and metal ions) on the direction of metabolic flow and the corresponding metabolic flux, a large accumulation of α-ketoglutarate is achieved. Background technique [0002] α-ketoglutaric acid, also known as α-gluconic acid, 2-oxoglutaric acid or α-oxoglutaric acid, is one of the important intermediate products in the tricarboxylic acid (TCA) cycle, and is found in microbial cells. It plays an important role in metabolism and is also an important precursor substance for the synthesis of various amino acids and proteins. Its structural formula is: [0003] [0004] Alpha-ketoglutarate structure [0005] α-Ketoglutaric acid has important application prospects in the fields of medicine, organic synthesis, and nutritional enhancers. At present, the main application ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/50C12R1/645
Inventor 陈坚张旦旦刘立明堵国成
Owner JIANGNAN UNIV
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