Functional roles and substrate specificities of twelve cytochromes P450 belonging to CYP52 family in n-alkane assimilating yeast Yarrowia lipolytica
Functional roles and substrate specificities of twelve cytochromes P450 belonging to CYP52 family in n-alkane assimilating yeast Yarrowia lipolytica
复制标题
DOI：
10.1016/j.fgb.2016.03.007
复制
发表时间：
2016-06-01
影响因子：
3
通讯作者：
Fukuda, Ryouichi
中科院分区：
文献类型：
--
作者：
Iwama, Ryo;Kobayashi, Satoshi;Fukuda, Ryouichi
解脂Yarrowia拥有12个ALK基因，编码细胞色素P450家族的细胞色素P450。本研究以一株alk基因全部缺失的解脂耶尔森菌为材料，构建了分别表达alk基因的菌株，并通过观察它们在正构烷烃上的生长情况和分析脂肪酸代谢来确定它们的作用和底物特异性。结果表明，根据底物的特异性，这12个ALK蛋白可以分为四类：Alk1p、Alk2p、Alk9p和Alk10p，它们具有显著的羟化正构烷烃的活性；Alk4p、Alk5p和Alk7p，它们具有显著的羟化十二烷酸共末端的活性；Alk3p和Alk6p，它们具有显著的羟化正构烷烃和十二酸的活性；以及Alk8p、Alk11p和Alk12p，它们对这些底物的氧化活性很弱或没有活性。通过测定12个ALK基因缺失突变株在含十二醇的培养基中的活力和观察突变株在十二醛上的生长情况，分析了ALK蛋白在脂肪醇和脂肪醛氧化反应中的作用。推测alk基因(S)参与了十二醇的解毒和十二醛的同化。这些结果表明，编码CYP52家族P450的基因在解脂耶尔森菌中发生了倍增和多样化，以同化各种疏水化合物。(C)2016 Elsevier Inc.保留所有权利。
Yarrowia lipolytica possesses twelve ALK genes, which encode cytochromes P450 in the CYP52 family. In this study, using a Y. lipolytica strain from which all twelve ALK genes had been deleted, strains individually expressing each of the ALK genes were constructed and their roles and substrate specificities were determined by observing their growth on n-alkanes and analyzing fatty acid metabolism. The results suggested that the twelve Alk proteins can be categorized into four groups based on their substrate specificity: Alk1p, Alk2p, Alk9p, and Alk10p, which have significant activities to hydroxylate n-alkanes; Alk4p, Alk5p, and Alk7p, which have significant activities to hydroxylate the co-terminal end of dodecanoic acid; Alk3p and Alk6p, which have significant activities to hydroxylate both n-alkanes and dodecanoic acid; and Alk8p, Alk11p, and Alk12p, which showed faint or no activities to oxidize these substrates. The involvement of Alk proteins in the oxidation of fatty alcohols and fatty aldehydes was also analyzed by measuring viability of the mutant deleted for twelve ALK genes in medium containing dodecanol and by observing growth on dodecanal of a mutant strain, in which twelve ALK genes were deleted along with four fatty aldehyde dehydrogenase genes. It was suggested that ALK gene(s) is/are involved in the detoxification of dodecanol and the assimilation of dodecanal. These results imply that genes encoding CYP52-family P450s have undergone multiplication and diversification in Y. lipolytica for assimilation of various hydrophobic compounds. (C) 2016 Elsevier Inc. All rights reserved.