Buffered glycerol-complex medium nsvub

The positively transformed yeast cells were cultured for about 20 h in a shaking flask comprising 50ml Buffered Glycerol Complex Medium (BMGY, 1% yeast extract,2% peptone, 100mM potassium phosphate buffer, pH 5.0, 1.34% YNB, 4 × 10-5 % biotin, and 1% glycerol) to OD 600 = 4.0.

BMGY medium (buffered glycerol complex medium) contained 10 g/liter Bacto yeast extract, 20 g/liter hipolypepton, 13.4 g/liter yeast nitrogen base without amino acids (YNB) (BD Biosciences), 0.4 mg/liter biotin (Nacalai Tesque), 100 mM potassium phosphate buffer (pH 6.0), and 20 g/liter glycerol. Oct 15, 2018 · The protein expressing strains were obtained using G418 selection and cultured in 25 ml of buffered glycerol-complex medium [BMGY; 1% yeast extract, 2% peptone, 100 mM potassium phosphate (pH 6.0 Oct 29, 2010 · Mut S phenotype integrated into the grown colonies with the AOX1 locus of the yeast genome were selected and grown in 1 L of buffered glycerol complex medium to an optical density over 6 at 600 nm. Cells were induced with 1% methanol by replacing the buffered glycerol complex medium with buffered methanol complex medium. Apr 29, 2003 · Protein expression was carried out at room temperature in a 96-well-plate format with buffered glycerol-complex medium (BMGY) consisting of 1% yeast extract, 2% peptone, 100 mM potassium phosphate buffer (pH 6.0), 1.34% yeast nitrogen base, 4 × 10 −5 % biotin, and 1% glycerol as a growth medium. The induction medium was buffered methanol A single colony was picked and expanded in buffered glycerol complex medium (BMGY) to create working cell banks (WCW). Inocula from WCW were grown in BMGY medium incubated at 30°C for fermentations in Basal Salt Medium (BSM) 25 or 25°C for fermentations in Rich Defined Medium (RDM), 26 250 rpm until they reached an OD 600 ~10. After growth for 48 h, the positive colonies were picked and seeded into buffered glycerol complex medium, followed by incubation at 30°C for 2 days. The resulting cultures were collected by centrifugation and resuspended for induction in buffered complex medium with 0.5% methanol.

BMGY medium (buffered glycerol complex medium) contained 10 g/liter Bacto yeast extract, 20 g/liter hipolypepton, 13.4 g/liter yeast nitrogen base without amino acids (YNB) (BD Biosciences), 0.4 mg/liter biotin (Nacalai Tesque), 100 mM potassium phosphate buffer (pH 6.0), and 20 g/liter glycerol.

Mar 05, 2019 · For efficient expression of recombinant protein, thawed freeze cultures were grown in a Buffered Glycerol-complex medium (BM* (70%, v/v), 0.5 M potassium buffer pH 6.0 (10%, v/v), YNB (1.4%, w/v Expression cell lines were cultured in BMGY (buffered glycerol complex) medium at room temperature (22°C) with shaking until the D 600 value reached 40–80. The BMGY growth medium was removed by centrifugation and replaced by BMMY (buffered minimum methanol) medium to induce the overexpression and secretion of YKL-39.

A 1 mL aliquot was then used to inoculate a 1 L culture of buffered glycerol complex medium [1% yeast extract, 2% peptone, 100 mM potassium phosphate (pH 6), 1.34% yeast nitrogen base, 4 × 10 −5 % biotin, 1% glycerol]. Cells were grown at 25°C to an optical density of 10.