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
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
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...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 