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Background   Pichia pastoris  is one of the most important host organisms for the recombinant production of proteins in industrial biotechnology. To date, strain specific parameters, which are needed to set up feeding profiles for fed batch cultivations, are determined by time-consuming continuous cultures or consecutive fed batch cultivations, operated at different parameter sets.    Results  Here, we developed a novel approach based on fast and easy to do batch cultivations with methanol pulses enabling a more rapid determination of the strain specific parameters specific substrate uptake rate q s , specific productivity q p  and the adaption time (Δtime adapt ) of the culture to methanol. Based on q s , an innovative feeding strategy to increase the productivity of a recombinant  Pichia pastoris  strain was developed. Higher specific substrate uptake rates resulted in increased specific productivity, which also showed a time dependent trajectory. A dynamic feeding strategy, where the setpoints for q s  were increased stepwise until a q s   max  of 2.0 mmol·g -1 ·h -1  resulted in the highest specific productivity of 11 U·g -1 ·h -1 .    Conclusions  Our strategy describes a novel and fast approach to determine strain specific parameters of a recombinant  Pichia pastoris  strain to set up feeding profiles solely based on the specific substrate uptake rate. This approach is generic and will allow application to other products and other hosts.

A dynamic method based on the specific substrate uptake rate to set up a feeding strategy for Pichia pastoris