Cloning and characterization of the peroxisomal acyl CoA oxidase ACO3 gene from the alkane-utilizing yeast Yarrowia lipolytica.
The ACO3 gene, which encodes one of the acyl-CoA oxidase isoenzymes, was isolated from the alkane-utilizing yeast Yarrowia lipolytica as a 10 kb genomic fragment. It was sequenced and found to encode a 701-amino acid protein very similar to other ACOs, 67.5% identical to Y. lipolytica Aco1p and about 40% identical to S. cerevisiae Pox1p. Haploid strains with a disrupted allele were able to grow on fatty acids. The levels of acyl-CoA oxidase activity in the ACO3 deleted strain, in an ACO1 deleted strain and in the wild-type strain, suggested that ACO3 encodes a short chain acyl-CoA oxidase isoenzyme. This narrow substrate spectrum was confirmed by expression of Aco3p in E. coli.
Wang
H
H
Laboratoire de Génétique des Microorganismes, CNRS ERS567, INRA Centre de Grignon, Thiverval Grignon, France.
Le Clainche
A
A
Le Dall
M T
MT
Wache
Y
Y
Pagot
Y
Y
Belin
J M
JM
Gaillardin
C
C
Nicaud
J M
JM
D042962:Acyl-CoA Oxidase; D000473:Alkanes / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D005800:Genes, Fungal; D008830:Microbodies / Q000201:enzymology; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D015183:Restriction Mapping; D004718:Saccharomycetales / Q000201:enzymology / Q000235:genetics*; D017422:Sequence Analysis, DNA
D000473:Alkanes; D010088:Oxidoreductases; D042962:Acyl-CoA Oxidase