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.

Feb 27, 2017 · Buffered glycerol-complex medium. CD: Circular dichroism. HPLC: High performance liquid chromatography. pNPR: p-Nitrophenyl α-L-rhamnopyranoside. YNB: Yeast nitrogen base. YPD: Yeast extract peptone dextrose medium Applications of the GST- Affinity Tag in the Purification and Characterization of Proteins A dissertation submitted in partial fulfillment of the requirements for the degree of May 23, 2017 · The transformants were selected on YPD (yeast extract peptone dextrose) plates containing 200 μg/mL zeocin. The selected clones were inoculated and amplified in 30 mL of buffered glycerol-complex medium at 30 °C for 2 days. Then the culture medium was replaced by 20 mL of buffered methanol-complex medium to induce protein expression. Twenty transformants were used to inoculate 40 ml of buffered glycerol complex medium (BMGY) (Invitrogen) (0.1 M potassium phosphate buffer at pH 6.0 containing 1% [wt/vol] yeast extract, 2% [wt/vol] peptone, 1.34% [wt/vol] yeast nitrogen base without amino acids, 1% [vol/vol] glycerol, and 4 × 10 −5 % [wt/vol] biotin). Cultures were grown Then, the culture was transferred into 1 L fresh BMGY, and all of the cells were collected by centrifugation at room temperature (3,500 g for 5 min) until the of the culture reached approximately 6, at which point it was resuspended in 100 mL of buffered methanol-complex medium (BMMY). Methanol (3%) was added to the media daily to induce the Jul 27, 2017 · The cells were then recovered by adding 500 µL liquid LB medium and incubating at 37 °C for 1 h and then plated on LB plates supplemented with 25 μg/mL Zeocin (TFS, R25001). After incubated at 37 °C overnight, ten colonies were cultured in 3 mL liquid LB medium supplemented with 25 µg/mL Zeocin at 37 °C overnight. The recombinants were cultured in Buffered Glycerol- complex Medium (BMGY) at 30℃ and then resuspended in Buffered Methanol-complex Medium (BMMY). The cultures were shaken at 250 r/min for 108 h at 29℃, and the methanol concentration retained 0.5% (w/v). Column chromatography The supernatant was harvested by 3600 rpm centrifugation

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.

The applications of viral protein cages have expanded rapidly into the fields of bionanotechnology and materials science. However, the low-cost produc… 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. 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

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. Methanol was added at a concentra-tion of 1% every 24 hours during the induction phase (up to 72 hours). Cultures were centrifuged

Dec 03, 2019 · Xylanase is one of the most extensively used biocatalysts for biomass degradation. However, its low catalytic efficiency and poor thermostability limit its applications. Therefore, improving the properties of xylanases to enable synergistic degradation of lignocellulosic biomass with cellulase is of considerable significance in the field of bioenergy. Using fragment replacement, we improved