Overexpression of Saccharomyces cerevisiae mannitol dehydrogenase gene (YEL070w) in glycerol synthesis-deficient S. Cerevisiae mutant

Expression of glycerol 3‐phosphate dehydrogenase gene (CvGPD1) in salt‐tolerant yeast Candida versatilis is stimulated by high concentrations of NaCl

“…Yeast C. versatilis NBR10650 (wild-type), S. cerevisiae YSH642 (Ansell et al, 1997), and W303-1A (Thomas and Rothstein, 1989;Wallis et al, 1989) were cultivated with shaking in YPAD medium (Watanabe et al, 2005) or Leu-dropout medium (Watanabe et al, 2006) after pre-cultivation for 3 days at 30 • C. To induce gene transcription, after precultivation in Leu-dropout medium containing 2% raffinose, pre-culture was added to Leu-dropout medium containing 1% raffinose and 1% galactose and cultivated with shaking at 30 • C. Escherichia coli JM109 (competent cells were purchased from Takara) was cultivated with shaking in LB medium (Sambrook and Russel, 2001) at 37 • C. Culture plates were prepared by solidifying liquid medium through the addition of 2% w/v agar and were incubated at 30 • C or 37 • C.…”

“…Measurement of GPDH enzyme activity, and glycerol and protein levels GPDH enzyme activity was measured as described previously, with some modification (Watanabe et al, 2006). Briefly, the reaction mixture consisted of 10 mM Tris-HCl buffer, pH 8.5, 100 µM snglycerol 3-phosphate (Sigma, G7886), 100 µM β-NAD + or β-NADP + and enzyme solution.…”