Open AccessRecombinant Protein Production with Pichia pastoris in Continuous Fermentation – Kinetic Analysis of Growth and Product Formation
Author(s) -
Curvers S.,
Linnemann J.,
Klauser T.,
Wandrey C.,
Takors R.
Publication year - 2002
Publication title -
engineering in life sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.547
H-Index - 57
eISSN - 1618-2863
pISSN - 1618-0240
DOI - 10.1002/1618-2863(20020806)2:8<229::aid-elsc229>3.0.co;2-9
Subject(s) - pichia pastoris , fermentation , methanol , yeast , pichia , recombinant dna , chemistry , substrate (aquarium) , biochemistry , growth rate , chromatography , biology , organic chemistry , gene , ecology , geometry , mathematics
Continuous fermentation was applied to the production of recombinant human chymotrypsinogen B (hCTRB) by the methylotrophic yeast Pichia pastoris as a tool for the kinetic analysis of growth and product formation. Using methanol as the sole source of carbon, energy, and induction, cell growth could be described by a non‐competitive Monod approach. Maximum growth rate μ max was determined to 0.084 h ‐‐1 and the K M ‐value for methanol to 0.22 g·L ‐‐1 , respectively. With respect to product formation, a similar model was established exhibiting a methanol concentration of 0.13 g·L ‐‐1 as the K M ‐value and a maximum biomass‐specific product‐formation rate of π max = 0.23 mg·g ‐‐1 ·h ‐‐1 . The production of hCTRB was strictly growth‐coupled. The data provided covers the range of methanol concentrations between 0 and 4 g·L ‐‐1 . Substrate concentrations exceeding this upper value led to a complete collapse of product formation. This change in phenotype turned out to be irreversible indicating a genetic instability of transformed Pichia pastoris caused by excess methanol.