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

Yarrowia lipolytica is a yeast which can utilize n-alkane as a sole carbon source. We isolated a Y. lipolytica peroxisomal acetoacetyl-CoA thiolase ge ne, 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 f eatures of peroxisomal acetoacetyl-CoA thiolase, We showed that the PAT1 di sruptant was not able to grow on n-decane, and that n-decane-inducible acet oacetyl-CoA thiolase activity largely depended on PAT1, The original mutant carried a mutation involving the replacement of Gly382 with Glu. This muta tion inactivated the ability of PAT1 to complement the defective n-decane u tilization of the disruptant, These results indicate that PAT1 encodes pero xisomal acetoacetyl-CoA thiolase and is essential for n-decane utilization in Y. lipolytica, (C) 2001 Academic Press.