CLONING AND CHARACTERIZATION OF THE PEROXISOMAL ACYL COA OXIDASE ACO3GENE FROM THE ALKANE-UTILIZING YEAST YARROWIA-LIPOLYTICA

The ACO3 gene, which encodes one of the acyl-CoA oxidase isoenzymes, w as isolated from the alkane-utilizing yeast Yarrowia lipolytica as a 1 0 kb genomic fragment. It was sequenced and found to encode a 701-amin o acid protein very similar to other ACOs, 67.5% identical to I: lipol ytica Acolp and about 40% identical to S. cerevisiae Poxlp. Haploid st rains with a disrupted allele were able to grow on fatty acids. The le vels of acyl-CoA oxidase activity in the ACO3 deleted strain, in an AC O1 deleted strain and in the wild-type strain, suggested that ACO3 enc odes a short chain acyl-CoA oxidase isoenzyme. This narrow substrate s pectrum was confirmed by expression of Aco3p in E. coli. (C) 1998 John Wiley & Sons, Ltd.