Method for producing l-tryptophan by bacterial fermentation with altered tdcd enzyme regulatory elements

A regulatory element and bacterial fermentation technology, which is applied in the field of amino acid fermentation, can solve the problems such as no attention has been paid to tdcD enzyme encoding gene, no tdcD enzyme regulatory element modification, etc., and achieve the effect of facilitating popularization and application and improving yield.

Active Publication Date: 2016-01-06
NINGXIA EPPEN BIOTECH
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Problems solved by technology

In the current literature reports, the attention through genetic engineering is mainly focused on mtr, aroP, tnaA, trpB, gnd, and traB In terms of isogenes (see Chinese patents 85101270, 93117586, 96111972, 200580043246, 200710056966, 201010598350, etc.), no attention has been paid to the tdcD enzyme and its coding gene for the production of L-tryptophan, let alone the modification of the regulatory elements of the tdcD enzyme , in fact the international patent WO2012135389 discloses the production of oxidation products of aromatic amino acids, which even requires the additional expression of heterologous tdc enzymes

Method used

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Examples

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Embodiment Construction

[0033] The content of the present invention is further illustrated below by way of examples. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art and commercially available common instruments and reagents, which can be found in "Molecular Cloning Experiment Guide (3rd Edition)" (Science Press), "Microbiological Experiments (4th Edition)" (Higher Education Press) and the manufacturer's instructions of the corresponding instruments and reagents, etc.

[0034] Construction Example Replaced with a transcriptionally less active promoter wxya upstream regulatory sequence

[0035] by Escherichia coli wxya We designed a weak transcriptional promoter (sequence shown in SEQIDNo: 2) and entrusted the Institute of Microbiology, Chinese Academy of Sciences to synthesize and construct it into the pMD-19T plasmid (available from Dalian Bao Biological Company) to replace the strain wxya The 337bp wild-type pro...

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Abstract

The invention provides a method for fermentatively producing L-tryptophan, which comprises transforming the bacterial chromosome wxya The wild-type regulatory element of the gene, so that the regulatory element of the transformed bacterium has its downstream enzyme gene (such as, wxya The ability to initiate expression of the gene) is weakened but not disappeared; and, L-tryptophan is produced by fermentation with the transformed bacteria. In addition, the present invention also provides methods and uses derived from this method, as well as polynucleotides, vectors and bacteria that can be used in these methods and applications.

Description

technical field [0001] The present invention belongs to the field of amino acid fermentation, in particular, the present invention relates to a method for fermentative production of l-tryptophan and its derivation method and application, as well as polynucleotides, vectors and bacteria that can be used in these methods and applications. Background technique [0002] Production of l-tryptophan by fermentation of l-tryptophan-producing bacteria (eg, Escherichia coli of the genus Escherichia and rod-shaped bacteria of the genus Corynebacterium) has been industrially applied. These bacteria can be bacteria isolated from nature, bacteria obtained through mutagenesis or genetic engineering, or both. In the current literature reports, the attention through genetic engineering is mainly focused on mtr, aroP, tnaA, trpB, gnd, and traB In terms of isogenes (see Chinese patents 85101270, 93117586, 96111972, 200580043246, 200710056966, 201010598350, etc.), no attention has been paid to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/22C12N15/70C12N1/21C12N15/11C12R1/19
Inventor 马吉银贾慧萍孟刚魏爱英马风勇李小刚杨立鹏
Owner NINGXIA EPPEN BIOTECH
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