A macrokinetic model-based comparative meta-analysis of recombinant protein production by Pichia pastoris under AOX1 promoter

Biotechnol Bioeng. 2015 Jun;112(6):1132-45. doi: 10.1002/bit.25518. Epub 2015 Apr 17.

Abstract

An unstructured macrokinetic model for heterologous production of Rhizopus oryzae lipase (ROL) by a Pichia pastoris PAOX1 based system was developed. Mean specific rates and state variables for various fed-batch cultures, under methanol limited and non-limited conditions were used for modeling. The most representative kinetic functions (viz., Monod, Haldane, Pirt and Luedeking-Piret) were used for the specific rates of cell growth (µ), substrate consumption (qS ) and product formation (qP ). The performance of two different process models was assessed via simulation tests, using an overall validation procedure. The best model describes the cellular growth by a non-monotonic substrate function, substrate uptake via Pirt's equation and product formation through a Luedeking-Piret equation. A comparative meta-analysis of heterologous protein production of various target proteins by Pichia pastoris under AOX1 promoter was conducted and a general strategy for improving protein production from process kinetics was developed as a key to bioprocess optimization.

Keywords: PAOX1; Pichia pastoris; bioprocess simulation; comparative analysis; recombinant protein production; unstructured macrokinetic model.

Publication types

  • Comparative Study
  • Meta-Analysis
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde Oxidase / genetics*
  • Culture Media / chemistry
  • Gene Expression*
  • Lipase / biosynthesis*
  • Lipase / genetics*
  • Pichia / genetics*
  • Pichia / growth & development
  • Pichia / metabolism*
  • Promoter Regions, Genetic*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Rhizopus / enzymology

Substances

  • Culture Media
  • Recombinant Proteins
  • Aldehyde Oxidase
  • Lipase