Isolation and characterization of polyhydroxyalkanoates inclusions and their associated proteins in Pseudomonas sp. 61-3

Biomacromolecules. 2002 Jul-Aug;3(4):787-92. doi: 10.1021/bm025516k.

Abstract

Two types of polyester inclusions of poly(3-hydroxybutyrate) [P(3HB)] and poly(3HB-co-3-hydroxyalkanoates) [P(3HB-co-3HA)] were isolated from crude extract of Pseudomonas sp. 61-3. Proteins associated with each inclusion were separated by SDS-PAGE. PHA synthase 1 (PhaC1(Ps)), PhaF(Ps), and PhaI(Ps) were identified from P(3HB-co-3HA) inclusions by N-terminal amino acid sequences analyses, as well as PHB synthase (PhbC(Ps)) and 24-kDa unknown protein were identified from P(3HB) inclusions. The structural genes of PhaF(Ps) and PhaI(Ps) were located downstream of the pha locus. The relative PHA/PHB synthase activities of each inclusion were measured for various 3-hydroxyacyl-coenzyme As of 4-12 carbon atoms. Direct atomic force microscopy observation of P(3HB) and P(3HB-co-3HA) inclusions demonstrated that the two types of inclusions had different morphologies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyltransferases / isolation & purification
  • Acyltransferases / metabolism
  • Bacterial Proteins / isolation & purification
  • Bacterial Proteins / metabolism
  • Blotting, Western
  • Inclusion Bodies / chemistry*
  • Inclusion Bodies / enzymology
  • Inclusion Bodies / ultrastructure
  • Microscopy, Atomic Force
  • Polyesters / chemical synthesis
  • Polyesters / isolation & purification*
  • Pseudomonas / chemistry

Substances

  • Bacterial Proteins
  • Polyesters
  • Acyltransferases
  • poly(3-hydroxyalkanoic acid) synthase