Wang H T, Rahaim P, Robbins P, Yocum R R
OmniGene, Inc., Cambridge, Massachusetts 02139-9002.
J Bacteriol. 1994 Nov;176(22):7091-5. doi: 10.1128/jb.176.22.7091-7095.1994.
The Saccharomyces diastaticus DAR1 gene was cloned by complementation in an Escherichia coli strain auxogrophic for glycerol-3-phosphate. DAR1 encodes an NADH-dependent dihydroxyacetone phosphate reductase (sn-glycerol-3-phosphate dehydrogenase [G3PDase; EC 1.1.1.8]) homologous to several other eukaryotic G3PDases. DAR1 is distinct from GUT2, which encodes a glucose-repressed mitochondrial G3PDase, but is identical to GPD1 from S. cerevisiae, a close relative of S. diastaticus. The level of DAR1-encoded G3PDase was increased about threefold in a medium of high osmolarity. Disruption of DAR1 in a haploid S. cerevisiae was not lethal but led to a decrease in cytoplasmic NADH-dependent G3PDase activity, an increase in osmotic sensitivity, and a 25% reduction in glycerol secretion from cells grown anaerobically on glucose.
通过在一株对3-磷酸甘油营养缺陷的大肠杆菌菌株中进行互补，克隆出了糖化酵母（Saccharomyces diastaticus）的DAR1基因。DAR1编码一种依赖NADH的磷酸二羟丙酮还原酶（sn-甘油-3-磷酸脱氢酶[G3PDase；EC 1.1.1.8]），与其他几种真核生物G3PDase同源。DAR1与GUT2不同，GUT2编码一种受葡萄糖抑制的线粒体G3PDase，但与糖化酵母的近亲酿酒酵母（S. cerevisiae）的GPD1相同。在高渗透压培养基中，DAR1编码的G3PDase水平增加了约三倍。单倍体酿酒酵母中DAR1的破坏并不致命，但导致细胞质中依赖NADH的G3PDase活性降低、渗透压敏感性增加，以及在葡萄糖上厌氧生长的细胞甘油分泌减少25%。
