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

Microbial Cell Factories

Research

Christoph Herwig¹  Christian Dietzsch¹  Oliver Spadiut¹ 

[1] Institute of Chemical Engineering, Research Area Biochemical Engineering, Vienna University of Technology, Vienna, Austria;

关键词: Feeding Strategy;    Substrate Uptake Rate;    High Specific Productivity;    Feeding Profile;    Yeast Nitrogen Base Medium;   

DOI  :  10.1186/1475-2859-10-14

 received in 2010-12-21, accepted in 2011-03-03,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundPichia 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.ResultsHere, 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 qs, specific productivity qp and the adaption time (Δtimeadapt) of the culture to methanol. Based on qs, 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 qs were increased stepwise until a qsmax of 2.0 mmol·g-1·h-1 resulted in the highest specific productivity of 11 U·g-1·h-1.ConclusionsOur 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.

【 授权许可】

Unknown   
© Dietzsch et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.