Enhanced induction of cytochromes P450alk that oxidize methyl-ends of n-alkanes and fatty acids in the long-chain dicarboxylic acid-hyperproducing mutant of Candida maltosa.
In the long-chain dicarboxylic acids (DCA)-hyperproducing mutant Candida maltosa strains, methyl-ends of n-alkanes and fatty acids are hydroxylated by n-alkane inducible cytochromes P450 (P450alk), presumably as an essential step in DCA production. A significantly higher production of P450alks was observed in response to n-alkane in the DCA-hyperproducing mutant strain M2030 than in the wild-type strain 1098. Northern analysis demonstrated that n-tetradecane induction levels of mRNAs of all four ALK genes encoding major P450alk isoforms involved in n-alkane assimilation were significantly higher in the DCA-hyperproducing mutant than in the wild-type strain. Among these four ALK genes, enhancement of the transcriptional induction level of ALK5, which prefers fatty acids as substrates, was prominent in the mutant. In agreement with Northern analysis, promoters of ALK genes, especially that of ALK5, more strongly responded to n-alkanes in the DCA-hyperproducing mutant than in the wild-type strain. These results suggest that the transcriptional control of ALK genes in the DCA-hyperproducing mutant strains was altered preferably to accelerate DCA production.
Kogure
Takahisa
T
Department of Biotechnology, The University of Tokyo, Bunkyo-ku, Tokyo, Japan. kogtaka@faculty.chiba-u.jp
Horiuchi
Hiroyuki
H
Matsuda
Hitoshi
H
Arie
Mami
M
Takagi
Masamichi
M
Ohta
Akinori
A
D000473:Alkanes / Q000494:pharmacology; D019677:Artificial Gene Fusion; D015152:Blotting, Northern; D015153:Blotting, Western; D002175:Candida / Q000235:genetics* / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000235:genetics; D003998:Dicarboxylic Acids / Q000378:metabolism*; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics; D005786:Gene Expression Regulation; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D006899:Mixed Function Oxygenases / Q000096:biosynthesis* / Q000235:genetics; D009154:Mutation; D011401:Promoter Regions, Genetic; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D012331:RNA, Fungal / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D000473:Alkanes; D003998:Dicarboxylic Acids; D005656:Fungal Proteins; D020033:Protein Isoforms; D012331:RNA, Fungal; D012333:RNA, Messenger; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C116330:CYP52A3 protein, Candida maltosa; D001616:beta-Galactosidase