Implications of evolutionary engineering for growth and recombinant protein production in methanol-based growth media in the yeast Pichia pastoris

Table 5 Results of fed batch cultivations of selected rHSA-expressing clones

Clone	YDM (g L⁻¹)	rHSA (mg L⁻¹)	Y_x/s glycerol (g g⁻¹)	Y_x/s methanol (g g⁻¹)	q_P (mg g⁻¹ h⁻¹)	Q_P (mg L⁻¹ h⁻¹)
X33 rHSA	82.2 ± 0.8	198.4 ± 0.4	0.666 ± 0.006	0.139 ± 0.004	0.009	1.39
Y250 3a rHSA	73.3 ± 0.8	34.2 ± 0.89	0.578 ± 0.011	0.093 ± 0.004	0.002	0.24
M250 1a rHSA	83.0 ± 0.6	151.7 ± 1.88	0.610 ± 0.003	0.141 ± 0.003	0.007	1.06
M250 3b rHSA	86.3 ± 0.1	350.0 ± 3.0	0.686 ± 0.011	0.149 ± 0.0	0.014	2.45

For cultivations a glycerol batch and fed batch were performed and MeOH pulses as well as constant feed phases were applied as described in the “Methods” section. YDM, rHSA yield and biomass yields (Y_x/s) are shown. Values represent averages ± standard deviation. Specific (q_P) and volumetric productivity (Q_P) were calculated for the entire methanol feed process as an average for all methanol fed batch and pulse phases. Biomass yield (Y_x/s) for glycerol was calculated for the glycerol batch phase. Biomass yield for methanol shows the average over all MeOH phases

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