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Construction and use for gentisic acid 1,2-dioxygenase gene engineering bacterium

A technology of genetically engineered bacteria and dioxygenase, applied in the direction of oxidoreductase, introduction of foreign genetic material by using vectors, recombinant DNA technology, etc., to achieve the effect of simple preparation method, convenient operation, suitable for large-scale batch production and application

Inactive Publication Date: 2005-03-16
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Abstract
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Problems solved by technology

On the one hand, maleic acid monoylpyruvate can be used as an analytical standard sample, and on the other hand, it may have potential uses (functional compounds, drug intermediates, etc.), but so far there is no supply of this compound on the market

Method used

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  • Construction and use for gentisic acid 1,2-dioxygenase gene engineering bacterium
  • Construction and use for gentisic acid 1,2-dioxygenase gene engineering bacterium

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Embodiment Construction

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Abstract

The invention relates to Klebsiella pneumoniae M5a1 gentisic acid 1, 2-bioxygenase gene engineered bacteria construction and uses thereof. Gentisic acid 1, 2-bioxygenase gene engineered bacteria construction method includes: LB medium ,engineered bacteria proliferation and following steps. (1)gentisic acid 1, 2-bioxygenase gene cloning and high expression (2) extraction of gentisic acid 1, 2-bioxygenase preparation of cisethylene dicarboxylic mono-acyl keto acetic acid using gentisic acid 1, 2-bioxygenase gene engineered bacteria. The invention is convenient to operate. Cisethylene dicarboxylic mono-acyl keto acetic acid product purity is high, being suitable for scale production and uses.

Description

technical field The invention relates to the construction and application of a gentisate 1,2-dioxygenase (gentisate 1,2-dioxygenase) genetically engineered bacterium from Klebsiella pneumoniae M5a1 (Klebsiella pneumoniae M5a1). Background technique Klebsiella pneumoniae M5a1 can degrade some aromatic compounds including hydroxybenzoic acid. Wherein, it is reported that this strain degrades 3-hydroxybenzoic acid into fumaric acid and pyruvate and enters the tricarboxylic acid cycle through the gentisic acid bypass pathway. The genes encoding each enzyme in this pathway are located on a DNA fragment about 8 kb in length digested by sphI. After sequencing analysis, the fragment contains eight reading frames, in which mhbD encodes gentisate 1,2-dioxygenase (gentisate 1,2-dioxygenase) to catalyze the conversion of gentisate into maleic acid monopyruvate (Malyylpyruvate, MP) (see Figure 2). On the one hand, maleyl aceruvate can be used as an analytical standard sample, and on ...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/70C12P7/50
Inventor 周宁一刘冬启高晓莉刘虹王淑君
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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