Figure 1.

Schematic representation of the methanol‐ or glucose‐based fed‐batch (FB) cultivations for secretome analysis of P. pastoris.

A. The processes comprised a glycerol batch phase, followed by a glycerol‐limited fed‐batch for 5 h. Then, methanol‐grown cultures received a methanol pulse, while glucose‐grown cultures were slowly fed with glucose to accumulate the same amount of biomass during this period (*). Afterwards, a constant methanol or glucose feed was applied for 67 h. See material and methods section for details of media composition and feed rates. Fed‐batch cultivations were performed in triplicates.

B. Sampling scheme for the analysis of the methanol and glucose fed‐batch processes. Samples were taken at the end of the glycerol batch phase (B1), the glycerol fed‐batch phase (G2), and at six time points during the methanol‐ or glucose‐limited fed‐batch phase (M3‐M8 and F3‐F8 respectively).