title: Two glycerol 3-phosphate dehydrogenase isogenes from Candida versatilis SN-18 play an important role in glycerol biosynthesis under osmotic stress.
abstract: Two isogenes of glycerol 3-phosphate dehydrogenase (GPD) from Candida versatilis SN-18 were cloned and sequenced. These intronless genes (Cagpd1 and Cagpd2) were both predicted to encode a 378 amino acid polypeptide, and the deduced amino acid sequences mutually showed 76% identity. Interestingly, Cagpd1 and Cagpd2 were located tandemly in a locus of genomic DNA within a 262 bp interval. To our knowledge, this represents a novel instance of isogenic genes relating to glucose metabolism. The stress response element (STRE) was found respectively at -93 to -89 bp upstream of the 5'end of Cagpd1 and -707 to -703 bp upstream of Cagpd2, indicating that these genes are involved in osmotic stress response. In heterologous expression using a gpd1Δgpd2Δ double deletion mutant of Saccharomyces cerevisiae, Cagpd1 and Cagpd2 transformants complemented the function of GPD, with Cagpd2 being much more effective than Cagpd1 in promoting growth and glycerol synthesis. Phylogenetic analysis of the amino acid sequences suggested that Cagpd1p and Cagpd2p are NADP(+)-dependent GPDs (EC 1.1.1.94). However, crude enzyme extract from Cagpd1 and Cagpd2 transformants showed GPD activity with only NAD(+) as cofactor. Hence, both Cagpd1p and Cagpd2p are likely NAD(+)-dependent GPDs (EC 1.1.1.8), similar to GPDs from S. cerevisiae and Candida magnoliae.
authors.affiliation: Laboratory of Applied Microbiology and Biotechnology, Department of Chemistry and Lifescience, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Daiki
authors.initials: D
authors.lastname: Mizushima
authors.affiliation: Laboratory of Applied Microbiology and Biotechnology, Department of Chemistry and Lifescience, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Iwata
authors.affiliation: Laboratory of Applied Microbiology and Biotechnology, Department of Chemistry and Lifescience, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Ishimaki
authors.affiliation: Laboratory of Applied Microbiology and Biotechnology, Department of Chemistry and Lifescience, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Ogihara
authors.affiliation: Laboratory of Applied Microbiology and Biotechnology, Department of Chemistry and Lifescience, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Kato
authors.affiliation: Laboratory of Applied Microbiology and Biotechnology, Department of Chemistry and Lifescience, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan. Electronic address: kasumi@brs.nihon-u.ac.jp.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Kasumi
mesh_terms: D000595:Amino Acid Sequence; D002175:Candida / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D005285:Fermentation; D005990:Glycerol / Q000378:metabolism*; D005993:Glycerolphosphate Dehydrogenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D005994:Glycerophosphates / Q000378:metabolism; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D009997:Osmotic Pressure; D010802:Phylogeny; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic
chemical_list: D005994:Glycerophosphates; D009243:NAD; D009249:NADP; C029620:alpha-glycerophosphoric acid; D005993:Glycerolphosphate Dehydrogenase; D005990:Glycerol
keywords: Candida; Fermentation; Glycerol; NAD(P)(+)-dependent glycerol 3-phosphate dehydrogenase; Osmotic condition