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Escherichia coli recombinant bacterium for efficiently utilizing fatty acid to synthesize glycine, and construction method and application thereof

A construction method and technology for recombinant Escherichia coli, applied in the field of Escherichia coli recombinant bacteria and its construction, can solve the problems of low enzyme activity, high cost of raw materials, and a large number of strains

Active Publication Date: 2019-01-25
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the above method can realize the production of glycine, there are problems of high raw material cost and low enzyme activity, and a large number of strains are required, so that biotransformation still needs to establish a cheaper raw material route

Method used

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  • Escherichia coli recombinant bacterium for efficiently utilizing fatty acid to synthesize glycine, and construction method and application thereof
  • Escherichia coli recombinant bacterium for efficiently utilizing fatty acid to synthesize glycine, and construction method and application thereof
  • Escherichia coli recombinant bacterium for efficiently utilizing fatty acid to synthesize glycine, and construction method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0052] Embodiment 1, construction of recombinant Escherichia coli engineering strains FG11, FG12, FG13, FG14, FG15, FG16, FG17

[0053] The construction of 7 recombinant Escherichia coli engineering strains FG11, FG12, FG13, FG14, FG15, FG16, FG17 includes the following twenty-six steps:

[0054] (1) Knockout of fatty acid degradation transcription factor fadR.

[0055] (1-a) First, prepare a P1 phage containing an Escherichia coli gene fragment, and the contained gene fragment has the trait of fadR knockout. The Escherichia coli gene fragment containing the fadR knockout trait comes from Escherichia coli strain JW1176, which is a W3110 series bacterial strain containing the fadR knockout trait, purchased from the National Institute of Genetics of Japan (NIG, Japan), wherein the fatty acid degradation transcription factor The gene fadR was replaced with a kanamycin resistance gene (about 1300bp) with FRT sites at both ends to knock out the fadR gene (Baba T, Ara T, et al. Con...

Embodiment 2

[0117] Embodiment 2, utilizing recombinant Escherichia coli strains FG11, FG12, FG13, FG14, FG15, FG16, FG17 to produce glycine with fatty acid as raw material

[0118] (27) A medium composition.

[0119] The solvent of medium A is water, and the solute and its final concentration are as follows:

[0120] NaHPO 4 : 25mM

[0121] K H 2 PO 4 : 25mM

[0122] NH 4 Cl: 50mM

[0123] Na 2 SO 4 : 5mM

[0124] MgSO 4 : 2mM

[0125] Glycerin: 0.5%

[0126] Yeast powder: 0.5%

[0127] Trace elements: 50μM FeCl 3 , 20 μM CaCl 2 , 10 μM MnCl 2 , 10 μM ZnSO 4 , 2 μM CoCl 2 , 2 μM NiCl 2 , 2 μM Na 2 MO 4 , 2 μM Na 2 SeO 3 and 2 μM H 3 BO 3 .

[0128] (28) B medium components.

[0129] The composition of medium B is based on the composition of medium A, and also contains 0.5% of palmitic acid and 0.2% of polyoxyethylene ether Brij58 emulsifier.

[0130] (29) C medium components.

[0131] C medium composition is based on A medium composition, and also contains pal...

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Abstract

The invention discloses an Escherichia coli recombinant bacterium for efficiently utilizing fatty acids to synthesize glycine and a construction method and application thereof. Escherichia coli recombinant bacterium contains one or more of that following genetic modification: (1) enhancement of fatty acid uptake pathway and fatty acid beta oxidation pathway; (2) glyoxylate cycle-related gene targets were modified to enhance glyoxylate synthesis; (3) introduction / enhancement of glutamate glyoxylate transaminase gene expression. The engineering strain of the invention has the ability to convertwhole cells of fatty acid substrates of different lengths into glycine, which can be stearic acid (C18), palmitic acid (C16), tetradecanoic acid (C14), lauric acid (C12), decanoic acid (C10), octanoicacid (C8) or caproic acid (C6). The ability of the engineered strain to synthesize glycine from fatty acids makes it useful for the industrial production of glycine products.

Description

technical field [0001] The invention relates to the fields of biotechnology and biochemical industry of industrial microorganisms, in particular to Escherichia coli recombinant bacteria that efficiently utilize fatty acids to synthesize glycine and its construction method and application. Background technique [0002] With the development of society, human beings are facing a series of severe challenges such as energy shortage, resource shortage, environmental degradation, economic recession and climate change, and new industries such as biomanufacturing and bioenergy are emerging. Compared with traditional chemical production, biotechnology has the advantages of energy saving, emission reduction and consumption reduction. Through genetic modification of industrial microbial strains, the utilization of raw materials by microbial cells can be enhanced, the conversion rate of products can be improved, and production costs can be reduced. [0003] Glycine (English name Glycine...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12P13/04C12R1/19
CPCC12P13/04C07K14/245
Inventor 刘伟丰向书漫刘波陶勇
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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