Fig. 2

Selection of optimal culture medium for rhBiP secretion in shake flask cultures of P. pastoris. SDS-PAGE was performed with following culture media samples on lanes: C- P. pastoris transformed with control pPIC3.5K vector in complex YEPM culture media; rhBiP-producing clone ppBiP10 cultivated in BSM – basal salt medium; BSM + DTT – basal salt medium supplemented with 2mM DTT; BSM + TCEP - basal salt medium supplemented with 2 mM TCEP; BSM + Gluc - basal salt medium supplemented with 0,75% (m/V) glucose; BSM + DTT + Gucose - basal salt medium supplemented with 2 mM DTT and 0.75% (m/V) glucose. All cultures (YEPM and BSM media with or without additives) contained the same 0.5% amount of the inducer methanol, as described in Methods. All culture growth media were concentrated 10x, and 7.5 µl (obtained from 75 µl of culture medium) was loaded onto the lane. M lane – unstained protein ladder (ThermoScientific, cat. no. 26614)