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Poly(hydroxyalkanoate) synthase genotype and PHA production of Pseudomonas corrugata and P. mediterranea

Daniel K Y Solaiman1, Vittoria Catara, Sebastiana Greco

  • 1Agricultural Research Service, US Department of Agriculture, Eastern Regional Research Center, 600 E Mermaid Lane, Wyndmoor, PA 19038, USA. dsolaiman@errc.ars.usda.gov

Journal of Industrial Microbiology & Biotechnology
|February 24, 2005
PubMed

Insights

Pseudomonas strains were analyzed for poly(hydroxyalkanoate) (PHA) synthase genes. This research distinguishes Pseudomonas corrugata strains into two groups and confirms PHA production in Pseudomonas mediterranea.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Pseudomonas corrugata and P. mediterranea are closely related bacterial species.
  • Poly(hydroxyalkanoates) (PHAs) are biopolyesters with diverse applications.
  • The genetic basis for PHA production, specifically the pha loci, can vary within bacterial species.

Purpose of the Study:

  • To investigate the presence and variability of PHA synthase genes (pha loci) in P. corrugata and P. mediterranea.
  • To characterize the phaC gene fragments and associated PHA composition in these Pseudomonas species.
  • To differentiate P. corrugata strains based on their pha loci and confirm PHA production in P. mediterranea.

Main Methods:

  • Polymerase Chain Reaction (PCR) was used to amplify segments of medium-chain-length PHA (mcl-PHA) synthase genes (phaC1 and phaC2).
  • Combined PCR-restriction endonuclease analysis and semi-nested PCR were employed for detailed detection.
  • Southern hybridization was performed on representative strains.
  • Nucleic acid sequencing and analysis of PHA composition (monomer analysis) were conducted.

Main Results:

  • All P. mediterranea strains possessed both phaC1 and phaC2 genes.
  • Six out of 56 P. corrugata strains contained both phaC1 and phaC2.
  • The remaining 50 P. corrugata strains showed only phaC2 via PCR, but Southern hybridization indicated the presence of phaC1.
  • Nucleic acid sequences of phaC fragments exhibited 95-100% similarity.
  • The predominant mcl-PHA monomers were beta-hydroxyoctanoate (33-47 mol%) and beta-hydroxydecanoate (26-36 mol%).

Conclusions:

  • The study successfully differentiated P. corrugata strains into two distinct groups based on pha loci.
  • This research provides the first evidence of mcl-PHA production in P. mediterranea.
  • The findings contribute to understanding the genetic diversity and biosynthetic capabilities of Pseudomonas species regarding PHAs.

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