Heterologous protein expression in the methylotrophic yeast Pichia pastoris

FEMS Microbiol Rev. 2000 Jan;24(1):45-66. doi: 10.1111/j.1574-6976.2000.tb00532.x.


During the past 15 years, the methylotrophic yeast Pichia pastoris has developed into a highly successful system for the production of a variety of heterologous proteins. The increasing popularity of this particular expression system can be attributed to several factors, most importantly: (1) the simplicity of techniques needed for the molecular genetic manipulation of P. pastoris and their similarity to those of Saccharomyces cerevisiae, one of the most well-characterized experimental systems in modern biology; (2) the ability of P. pastoris to produce foreign proteins at high levels, either intracellularly or extracellularly; (3) the capability of performing many eukaryotic post-translational modifications, such as glycosylation, disulfide bond formation and proteolytic processing; and (4) the availability of the expression system as a commercially available kit. In this paper, we review the P. pastoris expression system: how it was developed, how it works, and what proteins have been produced. We also describe new promoters and auxotrophic marker/host strain combinations which extend the usefulness of the system.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.
  • Review

MeSH terms

  • Animals
  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / genetics
  • Fungal Proteins / biosynthesis
  • Fungal Proteins / genetics
  • Gene Expression
  • Genetic Vectors
  • Glycosylation
  • Humans
  • Methanol / metabolism*
  • Mitochondrial Proteins
  • Oxidoreductases / genetics
  • Pichia / genetics
  • Pichia / growth & development
  • Pichia / metabolism*
  • Plant Proteins
  • Promoter Regions, Genetic / genetics
  • Protein Engineering
  • Protein Processing, Post-Translational / genetics
  • Proteins / genetics
  • Proteins / metabolism*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / metabolism
  • Viral Proteins / biosynthesis
  • Viral Proteins / genetics


  • Bacterial Proteins
  • Fungal Proteins
  • Mitochondrial Proteins
  • Plant Proteins
  • Proteins
  • Recombinant Proteins
  • Viral Proteins
  • Oxidoreductases
  • alternative oxidase
  • Methanol