来自利用烷烃的解脂耶氏酵母的过氧化物酶体酰基辅酶A氧化酶ACO3基因的克隆与特性分析
Cloning and characterization of the peroxisomal acyl CoA oxidase ACO3 gene from the alkane-utilizing yeast Yarrowia lipolytica.
作者信息
Wang H, Le Clainche A, Le Dall M T, Wache Y, Pagot Y, Belin J M, Gaillardin C, Nicaud J M
机构信息
Laboratoire de Génétique des Microorganismes, CNRS ERS567, INRA Centre de Grignon, Thiverval Grignon, France.
出版信息
Yeast. 1998 Nov;14(15):1373-86. doi: 10.1002/(SICI)1097-0061(199811)14:15<1373::AID-YEA332>3.0.CO;2-1.
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.
编码酰基辅酶A氧化酶同工酶之一的ACO3基因，作为一个10 kb的基因组片段，是从利用烷烃的解脂耶氏酵母中分离出来的。对其进行了测序，发现它编码一种701个氨基酸的蛋白质，与其他酰基辅酶A氧化酶非常相似，与解脂耶氏酵母Aco1p的同源性为67.5%，与酿酒酵母Pox1p的同源性约为40%。等位基因被破坏的单倍体菌株能够在脂肪酸上生长。ACO3缺失菌株、ACO1缺失菌株和野生型菌株中的酰基辅酶A氧化酶活性水平表明，ACO3编码一种短链酰基辅酶A氧化酶同工酶。通过在大肠杆菌中表达Aco3p证实了这种狭窄的底物谱。
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