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 μmax was determined to 0.084 h‐‐1 and the KM‐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 KM‐value and a maximum biomass‐specific product‐formation rate of πmax = 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.
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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