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Method for fermenting and producing l-tryptophan with bacteria with weakened tdcd enzyme expression and/or reduced enzyme activity

A technology of bacterial fermentation and enzyme activity, applied in the field of amino acid fermentation, to achieve the effect of facilitating application and increasing production

Active Publication Date: 2016-09-07
NINGXIA EPPEN BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 paid to the tdcD enzyme and its coding gene for the production of L-tryptophan. In fact, the international patent WO2012135389 discloses the production of aromatic amino acids Oxidation products, where even additional expression of heterologous tdc enzymes is required

Method used

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  • Method for fermenting and producing l-tryptophan with bacteria with weakened tdcd enzyme expression and/or reduced enzyme activity
  • Method for fermenting and producing l-tryptophan with bacteria with weakened tdcd enzyme expression and/or reduced enzyme activity

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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] Build the example so that wxya Gene expression inactivation and complete knockout

[0035] Using the pKD3 plasmid (available from Biovector, Inc) as a template, PCR amplification was performed with primers P1 and P2 (synthesized by Shanghai Yingjun Company), and a DNA fragment with a length of 1100 bp was obtained. Wherein, PCR was carried out as follows: denaturation at 94°C for 30 seconds, annealing at 52°C for 30 seconds, and extension at 72...

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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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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