Method for fermentation production of L-tryptophan by tdcD enzyme expression weakened and/or enzyme activity reduced bacteria

A technology of bacterial fermentation and tryptophan, which is applied in the field of amino acid fermentation to achieve the effect of increasing production and facilitating popularization and application

Active Publication Date: 2013-10-02
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 On isogenes (see Chinese patents 85101270, 93117586, 96111972, 200580043246, 200710056966, 201010598350, etc.), no attention has been

Method used

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  • Method for fermentation production of L-tryptophan by tdcD enzyme expression weakened and/or enzyme activity reduced bacteria
  • Method for fermentation production of L-tryptophan by tdcD enzyme expression weakened and/or enzyme activity reduced bacteria

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[0035] The content of the present invention is further illustrated by the following examples. Unless otherwise specified, the technical means used in the examples are conventional means known to those skilled in the art and commercially available instruments and reagents. For reference, please refer to "Molecular Cloning Experiment Guide (3rd Edition)" (Science Press), References such as "Microbiology Experiments (4th Edition)" (Higher Education Press) and the manufacturer's instructions for the corresponding instruments and reagents.

[0036]

[0037] Build the example to make tdcD Gene expression inactivation and complete knockout

[0038] Using the pKD3 plasmid (available from Biovector, Inc) as the template, PCR amplification was performed with primers P1 and P2 (commissioned by Shanghai Yingjun Company) to obtain a DNA fragment with a length of 1100 bp. Here, PCR was performed as follows: denaturation at 94°C for 30 seconds, annealing at 52°C for 30 seconds, and ext...

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Abstract

The invention provides a method for fermentation production of L-tryptophan. The method comprises: transforming bacteria to reduce their tdcD enzyme expression quantity and/or enzyme activity, even to enable their tdcD enzyme expression quantity and/or enzyme activity to completely disappear; and, applying the transformed bacteria to fermentation production of L-tryptophan. In addition, the invention also provides methods derived from the method and application, as well as polynucleotide, carriers and bacteria usable in the methods and application.

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 On isogenes (see Chinese patents 85101270, 93117586, 96111972, 200580043246, 200710056966, 201010598350, etc.), no attention has been paid to the tdcD...

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Application Information

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IPC IPC(8): C12P13/22C12N15/09C12N1/20C12N15/11C12R1/19
Inventor 马吉银魏爱英孟刚贾慧萍马风勇李小刚
Owner NINGXIA EPPEN BIOTECH
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