title: Functional roles and substrate specificities of twelve cytochromes P450 belonging to CYP52 family in n-alkane assimilating yeast Yarrowia lipolytica.
abstract: 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 ω-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.
authors.affiliation: Department of Biotechnology, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Iwama
authors.affiliation: Department of Biotechnology, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kobayashi
authors.affiliation: Department of Biotechnology, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Ishimaru
authors.affiliation: Department of Biological Chemistry, College of Bioscience and Biotechnology, Chubu University, 1200 Matsumoto-cho, Kasugai, Aichi 487-8501, Japan.
authors.forename: Akinori
authors.initials: A
authors.lastname: Ohta
authors.affiliation: Department of Biotechnology, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Horiuchi
authors.affiliation: Department of Biotechnology, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: afukuda@mail.ecc.u-tokyo.ac.jp.
authors.forename: Ryouichi
authors.initials: R
authors.lastname: Fukuda
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000235:genetics*; D000447:Aldehydes / Q000378:metabolism; D000473:Alkanes / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism; D005233:Fatty Alcohols / Q000378:metabolism; D017353:Gene Deletion; D010084:Oxidation-Reduction; D013379:Substrate Specificity; D025062:Yarrowia / Q000201:enzymology*
chemical_list: D000447:Aldehydes; D000473:Alkanes; D005227:Fatty Acids; D005233:Fatty Alcohols; C001634:fatty aldehyde; D003577:Cytochrome P-450 Enzyme System; D000444:Aldehyde Dehydrogenase
keywords: Cytochrome P450; Fatty acid; Yarrowia lipolytica; n-Alkane