Cloning and characterization of the peroxisomal acyl CoA oxidase ACO3 gene from the alkane-utilizing yeast Yarrowia lipolytica.

H. Wang , A. Le Clainche , M. T. Le Dall (1) , Yves Waché (2, 3, 4) , Y. Pagot (1) , J. M. Belin (3, 4) , C. Gaillardin (1, 5) , J. M. Nicaud (1, 5)

1 MGM - Microbiologie et Génétique Moléculaire
2 Microbiologie
3 GPMA - Génie des Procédés Microbiologiques et Alimentaires
4 Laboratoire de Microbiologie
5 Collection de Levures d'Intérêt Biotechnologique et Laboratoire de Génétique Moléculaire et Cellulaire

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.

Mots clés: Acyl-CoA Oxidase; Alkanes; Amino Acid Sequence; Base Sequence; Cloning; Escherichia coli; Gene Deletion; Genes; Fungal; Molecular Sequence Data; Oxidoreductases; Polymerase Chain Reaction; Restriction Mapping; Saccharomycetales; Sequence Analysis

Mots clés: Molecular; Microbodies; DNA

Domaines: Biotechnologie; Biochimie, Biologie Moléculaire

H. Wang, A. Le Clainche, M. T. Le Dall, Yves Waché, Y. Pagot, et al.. Cloning and characterization of the peroxisomal acyl CoA oxidase ACO3 gene from the alkane-utilizing yeast Yarrowia lipolytica.. Yeast, 1998, 14 (15), pp.1373-86. ⟨10.1002/(SICI)1097-0061(199811)14:15<1373::AID-YEA332>3.0.CO;2-1⟩. ⟨hal-00368240⟩