Isolation and characterization of acetoacetyl-CoA thiolase gene essential for n-decane assimilation in yeast Yarrowia lipolytica

Biochem Biophys Res Commun. 2001 Apr 6;282(3):832-8. doi: 10.1006/bbrc.2001.4653.

Abstract

Yarrowia lipolytica is a yeast which can utilize n-alkane as a sole carbon source. We isolated a Y. lipolytica peroxisomal acetoacetyl-CoA thiolase gene, PAT1, by complementation of a mutant that cannot utilize n-decane as a sole carbon source. We found that the putative PAT1 product had conserved features of peroxisomal acetoacetyl-CoA thiolase. We showed that the PAT1 disruptant was not able to grow on n-decane, and that n-decane-inducible acetoacetyl-CoA thiolase activity largely depended on PAT1. The original mutant carried a mutation involving the replacement of Gly382 with Glu. This mutation inactivated the ability of PAT1 to complement the defective n-decane utilization of the disruptant. These results indicate that PAT1 encodes peroxisomal acetoacetyl-CoA thiolase and is essential for n-decane utilization in Y. lipolytica.

MeSH terms

  • Acetyl-CoA C-Acetyltransferase / genetics*
  • Acetyl-CoA C-Acetyltransferase / metabolism*
  • Alkanes / metabolism*
  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers / genetics
  • Genes, Fungal*
  • Genetic Complementation Test
  • Genomic Library
  • Molecular Sequence Data
  • Mutation
  • Peroxisomes / enzymology
  • Saccharomycetales / enzymology*
  • Saccharomycetales / genetics*
  • Saccharomycetales / growth & development
  • Sequence Homology, Amino Acid

Substances

  • Alkanes
  • DNA Primers
  • Acetyl-CoA C-Acetyltransferase
  • decane

Associated data

  • GENBANK/AB042276