Gene capable of regulating and controlling inhibition of vitamin C fermented strain acid production during acid stress

An acid stress and gene technology, applied in fermentation, genetic engineering, plant genetic improvement, etc., can solve the problems of not being able to formulate metabolic engineering strategies and not understanding the microbial response mechanism.

Inactive Publication Date: 2013-09-18
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, rational and targeted metabolic engineering strategies cannot be developed without understanding the mechanism of microbial response to stress conditions.

Method used

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  • Gene capable of regulating and controlling inhibition of vitamin C fermented strain acid production during acid stress
  • Gene capable of regulating and controlling inhibition of vitamin C fermented strain acid production during acid stress
  • Gene capable of regulating and controlling inhibition of vitamin C fermented strain acid production during acid stress

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Differences in the Expression of Membrane Transporter Genes in Mixed Cultures and Individually Cultured Common Keto-Colonic Acid Bacteria

[0030] Table 1 Differences in the expression of membrane transporter genes in mixed bacteria culture and single culture

[0031]

[0032]

[0033]

[0034] As shown in Table 1, the expression levels of 45 membrane transporter genes in the common ketogenic cologronic acid bacteria cultured in the mixed culture were 2 times or more than that in the common ketogenic cologronic acid bacteria cultured alone, including 9 Amino acid transporter genes, 13 broad-spectrum ABC transporter genes, 11 oligopeptide transporter genes, 5 dipeptide transporter genes, 4 sulfur transporter genes, 2 metal ion transporter genes and 1 choline / Carnitine / betaine transporter genes (Table 2). From the above results, it can be seen that, compared with the common ketogenic cologne cultured alone, the common ketogenic cologne cells associate...

Embodiment 2

[0035] Example 2 Effects of acid stress treatment on the expression of the common ketogenic cologronic acid bacteria magic transporter gene expression in mixed culture and separate culture

[0036] Table 2 Differences in the expression of membrane transporter genes in the cells of V.

[0037]

[0038]

[0039]

[0040] Table 3 Differences in the expression of membrane transporter genes in the cells of Vulgaris vulgaris ketogenes alone cultured after acid stress treatment

[0041]

[0042]

[0043] After acid stress, 19 membrane transporter genes were up-regulated in the mixed cultured cells of V. , 6 iron ion transporter genes, 2 dipeptide transporter genes, 1 thiamine transporter gene and 1 MscS transporter gene (Table 4). In the common ketogenic cologne cultured alone, 22 membrane transporters were up-regulated after acid stress, including 7 amino acid transporter genes, 7 broad-spectrum ABC transporter genes, and 4 oligopeptide transporter genes. genes, 1 met...

Embodiment 3

[0057] Example 3 Adding iron ions inhibits acid production by fermentation of mixed bacteria

[0058] at 1m 3 Carried out in a fermenter, the concentration of iron ions in the initial fermentation medium is 1.59mg.L -1 , adding final concentrations of 2.38 (1.5 times), 3.18 (2 times) and 7.95 mg.L respectively -1 (5 times) iron ion, fermentation 72h. The results showed that the yield of 2-KLG decreased and the synthesis of 2-KLG was inhibited with the increase of iron ion concentration.

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Abstract

The invention relates to a gene capable of regulating and controlling inhibition of vitamin C fermented strain acid production during acid stress and belongs to the technical fields of basic research on vitamin C prepared by fermentation and biological engineering. The key gene for regulating and controlling yield reduction of acid stress induced 2-KLG in the production strains is determined, so a metabolic engineering strategy with high pertinence is generated. The expression condition of genes, in particular membrane transport protein genes, of specific microorganisms in the microorganisms subjected to mixed cultivation is analyzed by an expression profile chip technology. Compared with ketogenic gulonic acid bacteria subjected to single cultivation, the ketogenic gulonic acid bacteria subjected to mixed bacteria cultivation have 45 upregulated membrane transport protein genes and 15 downregulated membrane transport protein genes. The notable thing is that: 7 iron ion transport protein genes are inhibited to express. After acid stress, the iron ion transport protein genes are overexpressed. During fermentation, the iron ions are added to inhibit mixed bacteria to ferment and generate acid. The result shows that the iron ion transport protein exerts reverse action in the process of yield reduction of 2-KLG generated by the acid stress induced mixed bacteria fermentation.

Description

technical field [0001] The invention belongs to the technical field of basic research and bioengineering of vitamin C prepared by fermentation, and relates to a key gene for regulating the reduction of vitamin C output in fermentation strain cells during acid stress. Background technique [0002] Vitamin C, also known as ascorbic acid, is an essential vitamin and antioxidant for the human body. [0003] The vitamin C two-step fermentation process developed by my country with independent intellectual property rights is currently the only microbial transformation method successfully applied to the industrial production of vitamin C. The "two-step fermentation" process of vitamin C is as follows: the first step is to oxidize D-sorbitol to L-sorbose, commonly known as alcohol sugar conversion, and the second step is to further oxidize L-sorbose to the precursor of vitamin C2 -Keto-L-gulonic acid (2-keto-L-gulonic acid, 2-KLG), commonly known as sugar-acid conversion. The secon...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/02C12N15/10C12N15/31
Inventor 杨皓茹刘立明陈坚
Owner JIANGNAN UNIV
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