I wish to know the role of biotin in BMGY (Buffered Glycerol Complex Medium) Other articles mostly add it depending on the dissolved oxygen or methanol in the medium, but we do not have way to

ORIGINAL ARTICLE Cloning, characterization and heterologous expression of the first Penicillium echinulatum cellulase gene M.R. Rubini1, A.J.P. Dillon2, C.M. Kyaw1 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 Oct 02, 2012 · The P. pastoris transformants were cultured in 25 mL of buffered glycerol-complex medium (BMGY) shaken at 28°C and 250 rpm in 250-mL glass flasks. When cultures reached an OD 600 of about 6, the cells were centrifuged and resuspended in 100 mL of buffered methanol-complex medium (BMMY) to an OD 600 of 1.0, shaken at 28°C and 250 rpm in 500-mL After choosing the most resistant colon, a single colony of multiple inserted His −, Mut + GS115 recombinants was inoculated into 25 mL buffered glycerol complex medium (BMGY; 1% yeast extract, 2% peptone, 100 mM potassium phosphate (pH 6.0), 1.34% yeast nitrogen base (YNB), 4 × 10 - 5% biotin, 1% glycerol) and cultured at 250 rpm and 30°C The transformants were grown on minimal dextrose (MD) agar plates at 32°C for 48 h. Ninety-six colonies were randomly selected to grow in 2-mL buffered glycerol complex medium (BMGY) at 30°C for 48 h. The cells were collected by centrifugation (12,000g) and resuspended in 2-mL buffered methanol complex medium (BMMY). After 48-h induction with Buffered Glycerol Complex Medium, Sterile 2.0% Peptone, 1.0% Yeast extract, 1.34% Potassiumphosphate pH 6.0, 1.34% Yeast nitrogen base w/o AA, 0.4 mg/L Biotin, 1.0%

Jul 10, 2015 · For growth and induction, BMGY (buffered glycerol-complex medium) and BMMY (buffered methanol-complex medium), respectively, were used, both at pH 6.5 (EasySelectPichia expression kit; Invitrogen). P. pastoris transformants were screened for protein induction in 24-well plates as described ( Boettner et al., 2002 ).

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.

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. Dec 27, 2018 · Both enzymes were produced in shake‐flask cultivations, as previously described. 69 Briefly, precultures were grown in up to 750 mL of buffered glycerol‐complex medium (BMGY; 100 m m potassium phosphate buffer, pH 6.0, 2 % (w /v ) peptone, 1 % (w /v ) yeast extract, 4×10 −5 % (w /v ) biotin, 1 % (v /v ) glycerol) at 30 °C, 220 rpm. 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-complex medium (BMMY) consisting of 1.5% methanol instead of glycerol in BMGY. Minimal medium The resultant recombinant P. pastoris was inoculated in a 1-L Erlenmeyer flask with 200 mL of standard medium (1.34% [w/v] yeast nitrogen base, 2% [w/v] peptone, 1% [w/v] yeast extract, 0.4 mg/L biotin, and 100 mM potassium phosphate [pH 6.0]) containing 1% (w/v) glycerol (Buffered Glycerol-complex Medium). The inocula were placed in an