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 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.
Yamagami
S
S
Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
Iida
T
T
Nagata
Y
Y
Ohta
A
A
Takagi
M
M
D000101:Acetyl-CoA C-Acetyltransferase / Q000235:genetics* / Q000378:metabolism*; D000473:Alkanes / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D015698:Genomic Library; D008969:Molecular Sequence Data; D009154:Mutation; D020675:Peroxisomes / Q000201:enzymology; D004718:Saccharomycetales / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D017386:Sequence Homology, Amino Acid
D000473:Alkanes; D017931:DNA Primers; D000101:Acetyl-CoA C-Acetyltransferase; C012867:decane