Enhancement of cell viability and alkaline polygalacturonate lyase production by sorbitol co-feeding with methanol in Pichia pastoris fermentation

Bioresour Technol. 2010 Feb;101(4):1318-23. doi: 10.1016/j.biortech.2009.09.025. Epub 2009 Oct 1.

Abstract

Alkaline polygalacturonate lyase (PGL) production by Pichia pastoris GS115 was used as a model to study the mechanism and strategy for enhancing heterologous protein production. In order to enhance cell viability and volumetric recombinant protein productivity, sorbitol, which had been confirmed to be a non-repressive carbon source, was added together with methanol during the induction phase. The resultant PGL activity was up to 1593 U mL(-1), which was enhanced 1.85-fold compared to the control (863 U mL(-1)) cultured with sorbitol added at a constant rate of 3.6 g h(-1)L(-1) after an induction period of 100 h. Further results revealed that an appropriate sorbitol co-feeding strategy not only decreased the cell mortality to 8.8% (the control is about 23.1%) in the end of fermentation, but also reduced the proteolytic degradation of PGL.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Oxidoreductases / metabolism
  • Alkalies / metabolism
  • Biomass
  • Fermentation / drug effects*
  • Intracellular Space / drug effects
  • Intracellular Space / enzymology
  • Methanol / pharmacology*
  • Microbial Viability / drug effects*
  • Peptide Hydrolases / metabolism
  • Pichia / cytology*
  • Pichia / drug effects
  • Pichia / enzymology*
  • Polysaccharide-Lyases / biosynthesis*
  • Protein Processing, Post-Translational / drug effects
  • Sorbitol / pharmacology*
  • Temperature
  • Time Factors

Substances

  • Alkalies
  • Sorbitol
  • Alcohol Oxidoreductases
  • alcohol oxidase
  • Peptide Hydrolases
  • Polysaccharide-Lyases
  • pectate lyase
  • Methanol