Heterologous protein expression in the methylotrophic yeast Pichia pastoris.
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.
Cereghino
J L
JL
Department of Biochemistry and Molecular Biology, Oregon Graduate Institute of Science and Technology, 20000 N.W. Walker Road, Beaverton, OR, USA.
Cregg
J M
JM
D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006031:Glycosylation; D006801:Humans; D000432:Methanol / Q000378:metabolism*; D024101:Mitochondrial Proteins; D010088:Oxidoreductases / Q000235:genetics; D010843:Pichia / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D010940:Plant Proteins; D011401:Promoter Regions, Genetic / Q000235:genetics; D015202:Protein Engineering; D011499:Protein Processing, Post-Translational / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D000099188:Alternative Oxidase
D001426:Bacterial Proteins; D005656:Fungal Proteins; D000432:Methanol; D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D011506:Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D000099188:Alternative Oxidase