Production of recombinant proteins in fermenter cultures of the yeast Pichia pastoris

Current Opinion in Biotechnology

Volumn 13, Issue 4, 2002, Pages 329-332

  Cereghino, Geoff P Lin ^a   Cereghino, Joan Lin ^a   Ilgen, Christine ^b   Cregg, James M ^b  

^a UNIVERSITY OF THE PACIFIC   (United States)

^b KECK GRADUATE INSTITUTE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

FUNGAL PROTEIN; RECOMBINANT PROTEIN;

FERMENTATION TECHNIQUE; FUNGUS CULTURE; GENE EXPRESSION SYSTEM; PICHIA PASTORIS; PRIORITY JOURNAL; PROTEIN EXPRESSION; PROTEIN GLYCOSYLATION; PROTEIN PROCESSING; REVIEW;

FUNGI; PICHIA; PICHIA PASTORIS;

EID: 0036669602     PISSN: 09581669     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0958-1669(02)00330-0     Document Type: Review

References (28)

1. Higgins D.R., Cregg J.M., Methods in Molecular Biology: Pichia Protocols. 1998;Humana Press, Totowa, New Jersey. This book is the cookbook for manipulation of P. pastoris. Procedures ranging from strain construction (both classical- and molecular-genetic) to purification of recombinant proteins are described in detail. In particular, Chapter 9 by Stratton and colleagues provides a variety of detailed and tested procedures for culturing the yeast in fermenters. This is the place to start for expression with P. pastoris.
2. Lin Cereghino J., Cregg J.M. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev. 24:2001;45-66. A good introduction to P. pastoris as a heterologous expression system. The advantages and disadvantages of using P. pastoris for recombinant protein production are discussed. A list of over 300 proteins that have been successfully expressed in this yeast is provided along with their references.
3. Cregg J.M., Cereghino L., Shi J., Higgins D.R. Recombinant protein expression in Pichia pastoris. Mol Biotech. 16:2000;23-52.
4. Lin Cereghino G.P., Sunga A.J., Lin Cereghino J., Cregg J.M. Expression of foreign genes in the yeast Pichia pastoris. Setlow J.K., Genetic Engineering: Principles and Methods. 23:2001;157-169 Kluwer Academic/ Plenum Publishers, London. The review discusses the general steps involved in engineering P. pastoris to express a foreign protein. A discussion of some common problems and their solutions is provided, making this article a useful trouble-shooting guide.
5. Cregg J.M., Madden K.R. Development of the methylotrophic yeast, Pichia pastoris, as a host system for the production of foreign proteins. Dev Ind Microbiol. 29:1988;33-41.
6. Hellwig S., Emde F., Raven N.P.G., Henke M., van der Logt P., Fischer R. Analysis of single-chain antibody production in Pichia pastoris using on-line methanol control in fed-batch and mixed-feed fermentations. Biotech Bioeng. 74:2001;344-352. This paper demonstrates the inhibition of the AOX1 promoter by glycerol even at extremely low levels and shows the benefits of using a commercially available methanol sensor to monitor and adjust fermenter conditions so that optimized recombinant protein expression is achieved. The authors emphasize the benefits of an 'on-line' methanol control procedure used in mixed-feed conditions.
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9. Goodrick J.C., Xu M., Finnegan R., Schilling B.M., Schiavi S., Hoppe H., Wan N.C. High-level expression and stabilization of recombinant human chitinase produced in a continuous constitutive Pichia pastoris expression system. Biotech Eng. 76:2001;492-497. A continuous fermentation process is described in which the GAP promoter is used to constitutively express a recombinant human chitinase. This process obviates the need to use methanol for induction and has other benefits compared with traditional induction strategies.
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15. Callewaert N., Laroy W., Cadirgi H., Geysens S., Saelens X., Jou W.M., Contreras R. Use of HDEL-tagged Trichoderma reesei manosyl oligosaccharide 1,2-α-D-mannosidase for N-glycan engineering in Pichia pastoris. FEBS Lett. 503:2001;173-178. The authors describe their strategy to engineer a strain of P. pastoris that secretes proteins with more 'human-like' N-glycosylation. An HDEL retention signal is utilized to localize a recombinant mannosidase to the endoplasmic reticulum, where it can modify proteins in the secretory pathway, removing specific sugar groups. The authors detail how the engineered strain alters the glycosylation patterns on two different reporter glycoproteins, influenza virus haemagglutinin and Trypanosoma cruzi trans-sialidase. Although one protein is more efficiently altered than the other in terms of its expected glycosylation, the results raise hopes that N-glycan engineering can be accomplished in P. pastoris.
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21. Mochizuki S., Hamato N., Hirose M., Miyano K., Ohtni W., Kameyama S., Kuwae S., Tokuyama T., Ohi H. Expression and characterization of recombinant human antithrombin III in Pichia pastoris. Protein Expr Purif. 23:2001;55-65. The authors express, purify and characterize recombinant human antithrombin III. In depth biophysical analyses are used to assess the effect of O-glycosylation of several amino acid residues in relation to the antithrombin activity. O-Glycosylation in the reactive center loop of the protein is hypothesized to decrease the recombinant protein's inhibitory activity against thrombin.
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28. Werten M.W.T., Wisselink W.H., Jansen-van den Bosch T.J., de Bruin E.C., de Wolf F.A. Secreted production of a custom-designed, highly hydrophilic gelatin in Pichia pastoris. Protein Eng. 14:2001;447-454.