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Method of producing polyhydroxyalkanoates and special-purpose engineering bacterium for the same

A technology of polyhydroxyalkanoate and engineering bacteria, applied in the direction of bacteria, etc., can solve problems such as energy waste, high cost of oxygen consumption power, and metabolic imbalance

Inactive Publication Date: 2010-06-09
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

During this process, aerobic metabolism will also produce additional NADH, causing metabolic imbalance, resulting in high oxygen consumption during fermentation and waste of energy in raw materials
In addition, in the actual fermentation production process, the oxygen consumption power cost of the fermentation process is often as high as about 50% of the total energy consumption cost of fermentation.
Therefore, the existing PHA fermentation production technology has the problems of low conversion rate of carbon source and high fermentation cost.

Method used

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  • Method of producing polyhydroxyalkanoates and special-purpose engineering bacterium for the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1, utilize alcohol dehydrogenase promoter (being called for short P adhE ) and poly-3-hydroxybutyrate (PHB) synthesis related gene (phaCAB) to produce PHA and its effect verification

[0057] The method of following embodiment utilizes alcohol dehydrogenase promoter (being called for short P adhE ) and poly-3-hydroxybutyrate (PHB) synthesis-related gene (phaCAB) as an example to construct a recombinant vector pPadhE-CAB for hypoxia-induced expression of poly-3-hydroxybutyrate, and pPadhE-CAB was transformed by electroporation CAB introduced into wild Escherichia coli E.coli K-12 and its two strains of deletion mutant E.coli ackA of acetate synthesis pathway - and E. coli pta - Three hypoxia-induced engineered strains expressing poly-3-hydroxybutyrate were obtained. Wherein phaCAB is derived from plasmid pBHR68 (according to literature (Spiekermann, P., et al., A sensitive, viable-colony staining method using Nile red for direct screening of bacteria that a...

Embodiment 2

[0109] Example 2, Production of PHA under hypoxic conditions by using the promoter of pyruvate-formate lyase and phaCAB gene in series and its effect verification

[0110] The pyruvate-formate lyase promoter (referred to as P pflC ) and poly-3-hydroxybutyrate (PHB) synthesis-related gene (phaCAB) as an example to construct a recombinant vector pPpflC-CAB for hypoxia-induced expression of poly-3-hydroxybutyrate, and pPpflC-CAB was transformed by electroporation CAB introduced into wild Escherichia coli E.coli K-12 and its two strains of deletion mutant E.coli ackA of acetate synthesis pathway - and E. coli pta - Three hypoxia-induced engineering bacteria expressing poly-3-hydroxybutyrate (PHB) were obtained. Wherein phaCAB is derived from plasmid pBHR68 (according to literature (Spiekermann, P., et al., A sensitive, viable-colony staining method using Nile red fordirect screening of bacteria that accumulate polyhydroxyalkanoic acids and other lipid storage compounds. Arch Mic...

Embodiment 3

[0120] Embodiment 3, using the promoter of glycerol dehydrogenase (glycerol dehydrogenase is called for short GLD) gldA ) and phaC2 gene in series to produce PHA under hypoxic conditions

[0121] The following method utilizes the glycerol dehydrogenase promoter (referred to as P gldA ) and the low substrate specificity PHA synthesis gene (phaC2) (the 2873-4555th nucleotide sequence at the 5' end of GENBANK accession number AY278219) as an example to construct a recombinant vector for hypoxia-induced expression of poly-3-hydroxybutyrate pPgld-phaC2, pPgld-phaC2 was introduced into wild Escherichia coli E.coli K-12 and Ralstonia eutropha synthesis-deficient mutant strain Ralstonia eutropha PHB-4 by electroporation to obtain Escherichia coli engineering bacteria E.coii K -12 (pPgld-phaC2) or Roche eutropha engineering bacteria R.eutropha (pPgld-phaC2). Among them, the low substrate specificity PHA synthesis gene (phaC2) came from Pseudomonas stutzeri 1317 (Chen JY, Liu T, Zheng...

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Abstract

The invention discloses a method for producing polyhydroxy fatty acid ester and a specific project bacterial, which is a recombination bacterial after diverting recombination expression carrier with cascade anaerobic induced promoter and polyhydroxy fatty acid ester synthesis gene to host bacterial. The manufacturing method of polyhydroxy fatty acid ester is: getting polyhydroxy fatty acid ester by fermentation of the project bacterial in the anoxic condition. The invention doesn't need air or oxygen in the fermentation synthesis PHA process, which also doesn't need nitrogen, reduces the cost.

Description

technical field [0001] The invention relates to a method for producing polyhydroxyalkanoate and special engineering bacteria thereof. Background technique [0002] Polyhydroxyalkanoates (polyhydroxyalkanoates, referred to as PHA) is a class of polymer biopolyesters widely present in microorganisms, mainly as carbon sources and energy storage substances in organisms (Anderson AJ, Dawes EA. Occurrence, metabolism, metabolic role, and industrial use of bacterial polyhydroxyalkanoates. Microbiol. Rev., 1990, 54: 450-472; MadisonLL, Huisman GW. Metabolic engineering of poly(3-hydroxyalkanoates): from DNA to plastic. Microbiol. Mol. Biol. Rev., 1999 , 63:21-53). A variety of microorganisms in nature can synthesize PHA, and its molecular weight is generally from tens of thousands to several million (Sudesh K, Abe H, Doi Y.Synthesis, structure and properties of polyhydroxyalkanoates: biologicalpolyesters.Prog.Polym.Sci., 2000, 25 : 1503-1555). [0003] The general structural form...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12P7/64
Inventor 陈国强魏晓星吴琼
Owner TSINGHUA UNIV