Rearrangement of gene order in the phaCAB operon leads to effective production of ultrahigh-molecular-weight poly[(R)-3-hydroxybutyrate] in genetically engineered Escherichia coli.

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  • Author(s): Hiroe A;Hiroe A; Tsuge K; Nomura CT; Itaya M; Tsuge T
  • Source:
    Applied and environmental microbiology [Appl Environ Microbiol] 2012 May; Vol. 78 (9), pp. 3177-84. Date of Electronic Publication: 2012 Feb 17.
  • Publication Type:
    Journal Article; Research Support, Non-U.S. Gov't
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 7605801 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1098-5336 (Electronic) Linking ISSN: 00992240 NLM ISO Abbreviation: Appl Environ Microbiol Subsets: MEDLINE
    • Publication Information:
      Original Publication: Washington, American Society for Microbiology.
    • Subject Terms:
      Gene Order* ; Gene Rearrangement* ; Operon*; Escherichia coli/*genetics ; Escherichia coli/*metabolism ; Hydroxybutyrates/*metabolism ; Polyesters/*metabolism; Cupriavidus necator/genetics ; Cupriavidus necator/metabolism ; Gene Expression ; Genetic Vectors ; Hydroxybutyrates/chemistry ; Molecular Weight ; Organisms, Genetically Modified ; Plasmids ; Polyesters/chemistry ; Promoter Regions, Genetic
    • Abstract:
      Ultrahigh-molecular-weight poly[(R)-3-hydroxybutyrate] [UHMW-P(3HB)] synthesized by genetically engineered Escherichia coli is an environmentally friendly bioplastic material which can be processed into strong films or fibers. An operon of three genes (organized as phaCAB) encodes the essential proteins for the production of P(3HB) in the native producer, Ralstonia eutropha. The three genes of the phaCAB operon are phaC, which encodes the polyhydroxyalkanoate (PHA) synthase, phaA, which encodes a 3-ketothiolase, and phaB, which encodes an acetoacetyl coenzyme A (acetoacetyl-CoA) reductase. In this study, the effect of gene order of the phaCAB operon (phaABC, phaACB, phaBAC, phaBCA, phaCAB, and phaCBA) on an expression plasmid in genetically engineered E. coli was examined in order to determine the best organization to produce UHMW-P(3HB). The results showed that P(3HB) molecular weights and accumulation levels were both dependent on the order of the pha genes relative to the promoter. The most balanced production result was achieved in the strain harboring the phaBCA expression plasmid. In addition, analysis of expression levels and activity for P(3HB) biosynthesis enzymes and of P(3HB) molecular weight revealed that the concentration of active PHA synthase had a negative correlation with P(3HB) molecular weight and a positive correlation with cellular P(3HB) content. This result suggests that the level of P(3HB) synthase activity is a limiting factor for producing UHMW-P(3HB) and has a significant impact on P(3HB) production.
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    • Accession Number:
      0 (Hydroxybutyrates)
      0 (Polyesters)
      26063-00-3 (poly-beta-hydroxybutyrate)
    • Publication Date:
      Date Created: 20120221 Date Completed: 20120713 Latest Revision: 20220408
    • Publication Date:
      20240829
    • Accession Number:
      PMC3346486
    • Accession Number:
      10.1128/AEM.07715-11
    • Accession Number:
      22344649