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Recombinant Protein Production with Pichia pastoris in Continuous Fermentation – Kinetic Analysis of Growth and Product Formation

✍ Scribed by S. Curvers; J. Linnemann; T. Klauser; C. Wandrey; R. Takors

Publisher
John Wiley and Sons
Year
2001
Tongue
German
Weight
286 KB
Volume
73
Category
Article
ISSN
0009-286X

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✦ Synopsis

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 M O N O D approach. The maximum growth rate l max was determined to be 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 p 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.

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