Fermentation technique for improving yield of uridylic acid produced by saccharomyces cerevisiae engineered strain

A fermentation method, the technology of Saccharomyces cerevisiae, which is applied in the field of fermentation to improve the yield of uridine acid produced by Saccharomyces cerevisiae engineering bacteria, can solve a large number of foreign exchange, unfavorable nucleotide development, and limit the research and development of derivative products, etc., to improve the ability Effect

Inactive Publication Date: 2016-04-27
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most nucleotide products have been monopolized by Japanese companies for a long time. Domestic nucleotides even have to be exported to foreign countries at a low price, processed by foreign companies into final products and then sold back to China. This not only requires a large amount of foreign exchange, but also severely limits the research on corresponding derivative products. development, which is extremely unfavorable to the development of the domestic nucleotide market

Method used

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  • Fermentation technique for improving yield of uridylic acid produced by saccharomyces cerevisiae engineered strain
  • Fermentation technique for improving yield of uridylic acid produced by saccharomyces cerevisiae engineered strain
  • Fermentation technique for improving yield of uridylic acid produced by saccharomyces cerevisiae engineered strain

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Embodiment 1

[0026] Saccharomyces cerevisiae engineered strain pYX212-URA5-YPH499-URA6 - As an example, a fermentation method to increase the yield of uridine acid includes the following steps:

[0027] 1. Inoculate a ring of engineering bacteria into a centrifuge tube containing 6mL of YPD seed medium (yeast extract 10g / L, peptone 20g / L, glucose 20g / L), culture at 30°C and 200rpm for 24h, and then transfer to the tube containing In the Erlenmeyer flask of 100mL seed culture medium, 30 ℃, 200rpm cultivated for 24h, then transferred to 3L fresh wort medium (glucose 20g / L, yeast powder 10g / L, wort 400mL / L, Malt extract powder 10g / L, KH 2 PO 4 2.5g / L, MgSO 4 0.5g / L, FeSO 4 , 0.05g / L, urea 1.0g / L, adjust the pH to 5.0;) in the fermenter.

[0028] 2. Fermentation conditions: temperature at 30°C, ventilation rate at 2.5vvm, stirring speed at 400rpm, fermentation and cultivation for a certain period of time before entering the tank. The cells were collected by centrifugation, washed twice w...

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Abstract

The invention discloses a fermentation technique for improving yield of uridylic acid produced by a saccharomyces cerevisiae engineered strain. The fermentation technique comprises the following steps: preparing a malt extract medium; adding 1-5g of orotic acid, 0.1-0.3g of MgCl2, 1-2g of KH2PO4, 5-15g of glucose, 0.005-0.02g of manganese chloride, 1mL of glycerol, 0.5-2% (v / v) of methylbenzene and 15-25g of a yeast to 100mL of de-ionized water, regulating pH to 5.0, and reacting at 30 DEG C under 200rpm for 10h; and continuing to add 0.2-0.5mM of iodoacetic acid to the system, regulating temperature to 20 DEG C and pH value to 7.0, and ending the reaction system after reacting for 20-25h. By virtue of the method disclosed by the invention, the yield of the uridylic acid produced by the engineered strain is improved; and the method is applicable to the industries of functional food, health care products and medicines.

Description

technical field [0001] The invention belongs to the technical fields of food processing and medicine, and in particular relates to a method for improving the yield of uridine acid produced by Saccharomyces cerevisiae engineering bacteria (URA6 gene deletion and high-efficiency expression of orotate phosphoribosyltransferase and orotate nucleotide decarboxylase) fermentation method. Background technique [0002] Nucleotides, represented by uracil nucleotides, are extremely important biochemical substances in cell metabolism. In addition to being the precursors of DNA and RNA, they are also involved in cell growth and metabolism, energy storage and transformation, and immune responses. and signal transduction, etc. It plays an important role in helping to maintain the normal function of the body, improving cellular immunity, increasing antibody production, stimulating lymphocyte proliferation, and anti-oxidation. Uridine 5'-monophosphate (UMP) is a single nucleotide composed...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/30A23L33/00C12R1/865
CPCC12P19/305A23V2002/00A23V2250/61A23V2250/762
Inventor 杨雪莲王成涛张凯丽
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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