Studies on structure-function relationship of P450-multi-gene family from yeast and regulatory mechanism of their expression
Grant-in-Aid for Scientific Research (A)
|Allocation Type||Single-year Grants|
|Research Institution||The University of Tokyo|
TAKAGI Masamichi The University of Tokyo. Department of Biotechnology. Professor, 大学院・農学生命科学研究科, 教授 (50018339)
|Project Period (FY)
1996 – 1998
Completed(Fiscal Year 1998)
|Budget Amount *help
¥35,100,000 (Direct Cost : ¥35,100,000)
Fiscal Year 1998 : ¥5,700,000 (Direct Cost : ¥5,700,000)
Fiscal Year 1997 : ¥9,200,000 (Direct Cost : ¥9,200,000)
Fiscal Year 1996 : ¥20,200,000 (Direct Cost : ¥20,200,000)
|Keywords||Yeast / Candida maltosa / cytochrome P450 / CYP52 subfamily / substrate specificity / phylogenetic tree / n-alkane / fatty acid / hydroxylation / 氷酸化反応 / 遺伝子破壊 / 発現制御 / シスエレメント / 転写因子|
Some yeast species can assimilate n-alkane as a sole source of carbon and energy. We have been using Candida maltosa (an industrial microbe for dicarboxylic acid production) and Yarrowia lipolytica (a haploid-type yeast) to investigate the metabolism of n-alkane, regulation of gene expression and the mechanism of membrane proliferation in the cells when they are grown on n-alkane. We had already found that cytochromes P450, which are the primary oxigenases for n-alkane metabolism, are inducibly synthesized together with the proliferation of intracellular membrane where cytochromes P450 locate in the presence of n-alkane and that there are more than two genes for P450 in the genome of Candida maltosa. In the present investigation, we have analyzed the mechanisms of regulation of gene expression and of membrane proliferation in Candida maltosa and the gene structures for cytochromes P450 and the regulation of gene expression in Yarrowia lipolytica to understand the response of these cells at the molecular level to the most hydrophobic substrate, n-alkane. The results of our experiments consist of the following 6 chapters.
1. Isolation and characterization of several cytochromes P450 genes, and the substrate specificity of the encoded enzymes in Candida maltosa.
2. Analysis of the cis-elements involved in the regulation of expression of cytochromes P450 genes in Candida maltosa.
3. The system we have constructed to isolate genes for the trans-factors required for cytochrome P450 gene expression in Candida maltosa.
4. The mechanisms of intracellular membrane proliferation induced by n-alkane in Candida maltosa.
5. Isolation and characterization of several cytochromes P450 genes in Yarrowia lipolytica.
6. Some related experiments.
Research Products (19results)