RRC ID 36114
Author Mizushima D, Iwata H, Ishimaki Y, Ogihara J, Kato J, Kasumi T.
Title Two glycerol 3-phosphate dehydrogenase isogenes from Candida versatilis SN-18 play an important role in glycerol biosynthesis under osmotic stress.
Journal J. Biosci. Bioeng.
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.
Volume 121(5)
Pages 523-9
Published 2016-5
DOI 10.1016/j.jbiosc.2015.10.002
PII S1389-1723(15)00375-8
PMID 26906228
MeSH Amino Acid Sequence Candida / enzymology* Candida / genetics* Cloning, Molecular Fermentation Glycerol / metabolism* Glycerolphosphate Dehydrogenase / chemistry Glycerolphosphate Dehydrogenase / genetics* Glycerolphosphate Dehydrogenase / metabolism* Glycerophosphates / metabolism Molecular Sequence Data NAD / metabolism NADP / metabolism Osmotic Pressure* Phylogeny Saccharomyces cerevisiae / genetics Saccharomyces cerevisiae / growth & development Saccharomyces cerevisiae / metabolism Sequence Analysis, DNA Transformation, Genetic
IF 2.015
Times Cited 0
WOS Category BIOTECHNOLOGY & APPLIED MICROBIOLOGY FOOD SCIENCE & TECHNOLOGY
Resource
Yeast BYP148