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Method for producing 1,2-propanediol from recombinant Escherichia coli by the aid of glycerin

A technology of Escherichia coli and propylene glycol, applied in the field of recombinant Escherichia coli using glycerol to produce 1,2-propanediol, which can solve the problems of high synthesis cost, limited development of chemical synthesis methods, and difficulties in separation and purification

Inactive Publication Date: 2015-04-29
CHANGZHOU QIUHONG BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the synthesis of 1,2-propanediol by chemical synthesis, high temperature, high pressure and precious metal catalysts must be used
And there are many by-products, which brings difficulties to downstream separation and purification, and the synthesis cost is high, which limits the development of chemical synthesis methods

Method used

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  • Method for producing 1,2-propanediol from recombinant Escherichia coli by the aid of glycerin

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Embodiment 1

[0099] 1) Cloning of gene fragments

[0100]Using primer sequence 1 and primer sequence 2, the gene fragment mgsA was amplified, and the PCR amplification conditions were: 95°C for 30S, 62°C for 20S, 72°C for 30S, 30 cycles, and 72°C for 10 min. Using primer sequence 3 and primer sequence 4, the gene fragment gldA was amplified. The PCR amplification conditions were: 95° C. for 30 S, 62° C. for 20 S, 72° C. for 60 S, 30 cycles, and 72° C. for 10 min. Using primer sequence 5 and primer sequence 6, the gene fragment fucO was amplified. The PCR amplification conditions were: 95°C 30S, 62°C 20S, 72°C 60S, 30 cycles, and 72°C extension for 10 min.

[0101] 2) Construction of plasmid

[0102] The amplified gene fragment mgsA was digested with different restriction enzymes KpnI and SphI to obtain sticky ends, and the amplified gene fragment gldA was digested with different restriction enzymes SphI and SalI, To obtain sticky ends, the amplified gene fragment fucO was digested with d...

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Abstract

The invention relates to a method for producing 1,2-propanediol from recombinant Escherichia coli by the aid of glycerin. The method comprises steps as follows: 1) cloning genes mgsA, gldA and fucO involved in biosynthesis of the 1,2-propanediol; 2) connecting the genes with an Escherichia coli high-copy vector pUC19 to construct a recombinant vector of a coding gene containing protein required by biosynthesis of the 1,2-propanediol; 3) transferring a recombinant plasmid into the Escherichia coli BW25141, and obtaining a strain, which can realize the high yield of the 1,2-propanediol, of the Escherichia coli through selection marking by ampicillin; 4) producing the 1,2-propanediol through fermentation, and recovering the 1,2-propanediol from a fermentation liquid. The method has the advantages of environment-friendliness, high efficiency, stability, low cost and the like.

Description

Technical field: [0001] The invention belongs to the field of microbial genetic engineering. Specifically, three genes involved in the biosynthesis of 1,2-propanediol are cloned and connected to the high-copy vector pUC19 of Escherichia coli, the recombinant plasmid is transferred into Escherichia coli, and ampicillin is used as a screening marker , to enhance the expression of these three genes in Escherichia coli, so that 1,2-propanediol can be synthesized in a large amount in Escherichia coli. Background technique: [0002] 1,2-Propanediol (1,2-propanediol) is a colorless viscous hygroscopic liquid, miscible with water, ethanol and various organic solvents. The boiling point is 187.3°C. Melting point -60°C. The molecular formula is: C 3 h 8 o 2 , the structural formula is as follows. 1,2-propanediol is an important chemical raw material and intermediate, widely used in chemical industry, food, fuel and other fields, can be used as raw material for unsaturated polyes...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/70C12P7/18C12R1/19
CPCC12N9/0006C07K14/522C12N15/70C12P7/18C12Y101/01006
Inventor 刘立栋
Owner CHANGZHOU QIUHONG BIOTECH
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