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

Abstract：

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 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 qs 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.

journal_name

Microb Cell Fact

journal_title

Microbial cell factories

authors

Dietzsch C,Spadiut O,Herwig C

doi

10.1186/1475-2859-10-14

subject

Has Abstract

pub_date

2011-03-03 00:00:00

pages

14

issn

1475-2859

pii

1475-2859-10-14

journal_volume

10

pub_type

杂志文章

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