PHA synthase activity controls the molecular weight and polydispersity of polyhydroxybutyrate in vivo

Sang Jun Sim, Kristi D. Snell, Scott A. Hogan, Jo Anne Stubbe, Chokyun Rha, Anthony J. Sinskey

Research output: Contribution to journalArticle

154 Citations (Scopus)

Abstract

A synthetic operon for polyhydroxyalkanoate (PHA) biosynthesis designed to yield high levels of PHA synthase activity in vivo was constructed by positioning a genetic fragment encoding β-ketothiolase and acetoacetyl-CoA reductase behind a modified synthase gene containing an Escherichia coif promoter and ribosome binding site. Plasmids containing the synthetic operon and the native Alcaligenes eutro-phus PHA operon were transformed into E. coli DH5α and analyzed for polyhydroxybutyrate production. The molecular weight of polymer isolated from recombinant E. coli containing the modified synthase construct, determined by multiangle light scattering, was lower than that of the polymer from E. coli containing the natjve A. eutrophus operon. A further decrease in polyester molecular weight was observed with increased induction of the PHA biosynthetic genes in the synthetic operon. Comparison of the enzyme activity levels of PHA biosynthetic enzymes in a strain encoding the native operon with a strain possessing the synthetic operon indicates that the amount of polyhydroxyalkanoate synthase in a host organism plays a key role in controlling the molecular weight and the polydispersity of polymer.

Original languageEnglish
Pages (from-to)63-67
Number of pages5
JournalNature Biotechnology
Volume15
Issue number1
DOIs
Publication statusPublished - 1997 Jan

Keywords

  • Molecular weight
  • PHA synthase
  • Polyhydroxyalkanoates

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

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