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Method for ferment for producing adenomethionine with genetic engineering bacterium

A technology of adenosylmethionine and genetically engineered bacteria, which is applied in the field of fermenting and producing adenosylmethionine with genetically engineered bacteria, can solve the problems that the progress has not been reported, and achieve the effects of short cycle, low cost, and less pollution

Inactive Publication Date: 2008-05-07
WUHAN GRAND HOYO
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0013] The production of SAM by fermentation is still in its infancy in China, and the specific progress has not been reported. Fermentation production and extraction and refining technology are the main problems that need to be overcome in large-scale production

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  • Method for ferment for producing adenomethionine with genetic engineering bacterium

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Embodiment Construction

[0025]According to the SAM synthetase 2 gene (registration M23368) derived from S. cerevisiae in Genebank, the present invention designs two specific primers and obtains the enzyme gene through PCR amplification. The cloned SAM synthetase 2 gene derived from Saccharomyces cerevisiae was assembled into the expression vector pPIC9k of Saccharomyces cerevisiae to obtain the expression plasmid pPIC9k-SAM 2 , so that the SAM synthetase 2 gene is placed under the control of the promoter FLD1.

[0026] After the plasmids obtained above were linearized, they were transformed into the methanolic yeast Pichia pastoris host cell GS115 by means of protoplasts or electrotransformation. The promoter series or 3-terminal sequence in the expression plasmid improves homologous recombination with Pichi pastoris chromosomal DNA, so that the exogenous SAM synthetase 2 gene is integrated into the yeast chromosome and stably expressed in the recombinant cell. This recombinant expression mode integ...

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Abstract

The invention discloses a method fermentable producing adenosylmethionine utilizing genetic engineering bacteria, which is characterized in that the engineering bacteria is inoculated to a seed culture medium after activated in the plate culture medium, and then the material is inoculated to the fermentation culture medium for not less than one step after inoculated in the seed culture medium; glycerol is renewed as carbon source, obtaining the wet cells with a high density of 300 to 350g / L; then methanol and L-methionine are renewed for induction and transformation cultivation, obtaining the adenosylmethionine. The engineering bacteria is pichia pastoris GS115-sam II-208, and the book reservation number is CCTCC NO: M206106. The invention has the advantages of simple method, short cycle, high yield and low cost.

Description

technical field [0001] The invention belongs to biological fermentation engineering technology, in particular to a method for producing adenosylmethionine by fermenting genetically engineered bacteria. Background technique [0002] S-adenosylmethionine, also known as S-adenosyl-L-methionine, English name: S-adenosyl-L-methionine, chemical name: 5'-{[3S-3-amino-3-carboxy-propyl]- Methyl-S-sulfone}-5'-deoxyadenosine, commonly used abbreviations: SAM, SAMe, AdoMet, molecular formula: C 16 h 22 N 6 o 6 S. SAM is the active form of methionine, discovered by Cantoni in 1952. It is an important intermediate metabolite in organisms and participates in various biochemical reactions. In organisms, it is enzymatically synthesized from the substrate L-methionine and ATP by SAM synthetase (S-adenosylmethionine synthetase, ie ATP: L-methionineS-adenosyltransferase, EC2.5.1.6). SAM is a two-chiral substance with two isomers: (R, S)-SAM and (S, S)-SAM. Only (S,S)-SAM, ie (-)-SAM, ha...

Claims

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Application Information

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IPC IPC(8): C12P19/32C12N1/19C12N15/31
Inventor 刘胜祥
Owner WUHAN GRAND HOYO
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