Skip to main content
SpringerLink
Log in

Recombinant protein expression in Pichia pastoris

  • Review
  • Published:
Molecular Biotechnology Aims and scope Submit manuscript

Abstract

The methylotrophic yeast Pichia pastoris is now one of the standard tools used in molecular biology for the generation of recombinant protein. P. pastoris has demonstrated its most powerful success as a large-scale (fermentation) recombinant protein production tool. What began more than 20 years ago as a program to convert abundant methanol to a protein source for animal feed has been developed into what is today two important biological tools: a model eukaryote used in cell biology research and a recombinant protein production system. To date well over 200 heterologous proteins have been expressed in P. pastoris. Significant advances in the development of new strains and vectors, improved techniques, and the commercial availability of these tools coupled with a better understanding of the biology of Pichia species have led to this microbe’s value and power in commercial and research labs alike.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Romanos, M. A., Scorer, C. A., and Clare, J. J. (1992) Foreign gene expression in yeast: a review. Yeast 8, 423–488.

    Article  PubMed  CAS  Google Scholar 

  2. Cregg, J. M., Vedvick, T. S., and Raschke, W. C. (1993) Recent advances in the expression of foreign genes in Pichia pastoris. Biotechnology (NY) 11, 905–910.

    Article  CAS  Google Scholar 

  3. Romanos, M. (1995) Advances in the use of Pichia pastoris for high-level expression. Curr. Opin. Biotechnol. 6, 527–533.

    Article  CAS  Google Scholar 

  4. Cregg, J. M. (1999) Expression in the methylotrophic yeast Pichia pastoris. in Gene Expression Systems: Using Nature for the Art of Expression (Fernandez, J. M. and Hoeffler, J. P., eds.), Academic Press, San Diego, CA, pp. 157–191.

    Google Scholar 

  5. Cregg, J. M. and Higgins, D. R. (1995) Production of foreign proteins in the yeast Pichia pastoris. Can. J. Bot. 73(Suppl. 1), S981-S987.

    Google Scholar 

  6. Sreekrishna, K., Brankamp, R. G., Kropp, K. E., Blankenship, D. T., Tsay, J. T., Smith, P. L., Wierschke, J. D., Subramaniam, A., and Birkenberger, L. A. (1997) Strategies for optimal synthesis and secretion of heterologous proteins in the methylotrophic yeast Pichia pastoris. Gene 190, 55–62.

    Article  PubMed  CAS  Google Scholar 

  7. Gellissen, G. and Hollenberg, C. P. (1997) Application of yeasts in gene expression studies: a comparison of Saccharomyces cerevisiae, Hansenula polymorpha and Kluyveromyces lactis—a review. Gene 190, 87–97.

    Article  PubMed  CAS  Google Scholar 

  8. Higgins, D. R. (1995) Overview of protein expression in Pichia pastoris. In Current Protocols in Protein Science, Supplement 2 (Wingfield, P. T., ed.), John Wiley and Sons, New York, pp. 5.7.1–5.7.16.

    Google Scholar 

  9. Sreekrishna, K. (1993) Strategies for optimizing protein expression and secretion in the methylotrophic yeast Pichia pastoris. In Industrial Microorganisms: Basic and Applied Molecular Genetics (Baltz, R. H., Hegeman, G. D., and Skatrud, P. L., ed.), American Society for Microbiology, Washington, DC, pp. 119–126.

    Google Scholar 

  10. Higgins, D. R. and Cregg, J. M. (1998) Pichia Protocols, Methods in Molecular Biology, vol. 103, Humana Press, Totowa, NJ.

    Google Scholar 

  11. Cereghino, J. L. and Cregg, J. M. (2000) Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol. Rev., 24, 45–66.

    Article  PubMed  CAS  Google Scholar 

  12. Ogata, K., Nishikawa, H., and Ohsugi, M. (1969) A yeast capable of utilizing methanol. Agric. Biol. Chem. 33, 1519–1520.

    CAS  Google Scholar 

  13. Wegner, G. H. (1990) Emerging applications of the methylotrophic yeasts. FEMS Microbiol. Rev. 7, 279–283.

    PubMed  CAS  Google Scholar 

  14. Ellis, S. B., Brust, P. F., Koutz, P. J., Waters, A. F., Harpold, M. M., and Gingeras, T. R. (1985) Isolation of alcohol oxidase and two other methanol regulatable genes from the yeast Pichia pastoris. Mol. Cell. Biol. 5, 1111–1121.

    PubMed  CAS  Google Scholar 

  15. Cregg, J. M., Barringer, K. J., Hessler, A. Y., and Madden, K. R. (1985) Pichia pastoris as a host system for transformations. Mol. Cell. Biol. 5, 3376–3385.

    PubMed  CAS  Google Scholar 

  16. Tschopp, J. F., Brust, P. F., Cregg, J. M., Stillman, C. A., and Gingeras, T. R. (1987) Expression of the LacZ gene from two methanol-regulated promoters in Pichia pastoris. Nucleic Acids Res. 15, 3859–3876.

    Article  PubMed  CAS  Google Scholar 

  17. Cregg, J. M. and Madden, K. R. (1987) Development of yeast transformation systems and construction of methanol-utilization-defective mutants of Pichia pastoris gene disruption. In Biological Research on Yeasts, vol. 2 (Stewart, G. G., Russell, I., Klein, R. D., and Hiebsch, R. R., ed.), CRC Press, Boca Raton, FL, pp. 1–18.

    Google Scholar 

  18. Cregg, J. M., Madden, K. R., Barringer, K. J., Thill, G. P., and Stillman, C. A. (1989) Functional characterization of the two alcohol oxidase genes from the yeast Pichia pastoris. Mol. Cell. Biol. 9, 1316–1323.

    PubMed  CAS  Google Scholar 

  19. Koutz, P. J., Davis, G. R., Stillman, C., Barringer, K., Cregg, J. M., and Thill, G. (1989) Structural comparison of the Pichia pastoris alcohol oxidase genes. Yeast 5, 167–177.

    Article  PubMed  CAS  Google Scholar 

  20. Kurtzman, C. P. and Robnett, C. J. (1998) Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. Antonie van Leeuwenhoek 73, 331–371.

    Article  PubMed  CAS  Google Scholar 

  21. Veenhuis, M., van Dijken, J. P., and Harder, W. (1983) The significance of peroxisomes in the metabolism of one-carbon compounds in yeasts. Adv. Microb. Physiol. 24, 1–82.

    PubMed  CAS  Google Scholar 

  22. Couderc, R. and Baratti, J. (1980) Oxidation of methanol by the yeast Pichia pastoris: purification and properties of alcohol oxidase. Agric. Biol. Chem. 44, 2279–2289.

    CAS  Google Scholar 

  23. Tschopp, J. F., Sverlow, G., Kosson, R., Craig, W., and Grinna, L. (1987) High level secretion of glycosylated invertase in the methylotrophic yeast Pichia pastoris. Biotechnology (NY) 5, 1305–1308.

    Article  CAS  Google Scholar 

  24. Barr, K. A., Hopkins, S. A., and Sreekrishna, K. (1992) Protocol for efficient secretion of HSA developed from Pichia pastoris. Pharm. Eng. 12, 48–51.

    Google Scholar 

  25. Cregg, J. M., Tschopp, J. F., Stillman, C., Siegel, R., Akong, M., Craig, W. S., Buckholz, R. G., Madden, K. R., Kellaris, P. A., Davis, G. R., Smiley, B. L., Cruze, J., Torregrossa, R., Velicelebi, G., and Thill, G. P. (1987) High-level expression and efficient assembly of hepatitis B surface antigen in the methylotrophic yeast, Pichia pastoris. Biotechnology (NY) 5, 479–485.

    Article  CAS  Google Scholar 

  26. Chirulova, V., Cregg, J. M., and Meagher, M. M. (1997) Recombinant protein production in an alcohol oxidase-defective strain of Pichia pastoris in fedbatch fermentations. Enzyme Microb. Technol. 21, 277–283.

    Article  Google Scholar 

  27. Laroche, Y., Storme, V., De Muetter, J., Messens, J., and Lauwereys, M. (1994) High-level secretion and very efficient isotopic labeling of tick anticoagulant peptide (TAP) expressed in the methylotrophic yeast Pichia pastoris. Biotechnology (NY) 12, 1119–1124.

    Article  CAS  Google Scholar 

  28. Clare, J. J., Romanos, M. A., Rayment, F. B., Rowedder, J. E., Smith, M. A., Payne, M. M., Sreekrishna, K., and Henwood, C. A. (1991) Production of mouse epidermal growth factor in yeast: high-level secretion using Pichia pastoris strains containing multiple gene copies. Gene 105, 205–212.

    Article  PubMed  CAS  Google Scholar 

  29. Waterham, H. R., Digan, M. E., Koutz, P. J., Lair, S. V., and Cregg, J. M. (1997) Isolation of the Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase genc and regulation and use of its promoter. Gene 186, 37–44.

    Article  PubMed  CAS  Google Scholar 

  30. Shen, S., Sulter, G., Jeffries, T. W., and Cregg, J. M. (1998) A strong nitrogen source-regulated promoter for controlled expression of foreign genes in the yeast Pichia pastoris. Gene 216, 93–102.

    Article  PubMed  CAS  Google Scholar 

  31. Sreekrishna, K., Nelles, L., Potenz, R., Cruze, J., Mazzaferro, P., Fish, W., Fuke, M., Holden, K., Phelps, D., Wood, P., and Parker, K. (1989) High-level expression, purification, and characterization of recombinant human tumor necrosis factor synthesized in the methylotrophic yeast Pichia pastoris. Biochemistry 28, 4117–4125.

    Article  PubMed  CAS  Google Scholar 

  32. Scorer, C. A., Clare, J. J., McCombie, W. R., Romanos, M. A., and Sreekrishna, K. (1994) Rapid selection using G418 of high copy number transformants of Pichia pastoris for high-level foreign gene expression. Biotechnology (NY) 12, 181–184.

    Article  CAS  Google Scholar 

  33. Clare, J. J., Rayment, F. B., Ballantine, S. P., Sreekrishna, K., and Romanos, M. A. (1991) High-level expression of tetanus toxin fragment C in Pichia pastoris strains containing multiple tandem integrations of the gene. Biotechnology (NY) 9, 455–460.

    Article  CAS  Google Scholar 

  34. Romanos, M. A., Clare, J. J., Beesley, K. M., Rayment, F. B., Ballantine, S. P., Makoff, A. J., Dougan, G., Fairweather, N. F., and Charles, I. G. (1991) Recombinant Bordetella pertussis pertactin (P69) from the yeast Pichia pastoris: high-level production and immunological properties. Vaccine 9, 901–906.

    Article  PubMed  CAS  Google Scholar 

  35. Wung, J. L. and Gascoigne, N. R. (1996) Antibody screening for secreted proteins expressed in Pichia pastoris. Biotechniques 21, 808, 810, 812.

    PubMed  CAS  Google Scholar 

  36. Trimble, R. B., Atkinson, P. H., Tschopp, J. F., Townsend, R. R., and Maley, F. (1991) Structure of oligosaccharides on Saccharomyces SUC2 invertase secreted by the methylotrophic yeast Pichia pastoris. J. Biol. Chem. 266, 22807–22817.

    PubMed  CAS  Google Scholar 

  37. Gronwald, W., Loewen, M. C., Lix, B., Daugulis, A. J., Sonnichsen, F. D., Davies, P. L. and Sykes, B. D. (1998) The solution structure of type II antifreeze protein reveals a new member of the lectin family. Biochemistry 37, 4712–4721.

    Article  PubMed  CAS  Google Scholar 

  38. Kirkitadze, M. D., Krych, M., Uhrin, D., Dryden, D. T., Smith, B. O., Cooper, A., Wang, X., Hauhart, R., Atkinson, J. P., and Barlow, P. N. (1999) Independently melting modules and highly structured intermodular junctions within complement receptor type 1. Biochemistry 38, 7019–7031.

    Article  PubMed  CAS  Google Scholar 

  39. McAlister, M. S., Brown, M. H., Willis, A. C., Rudd, P. M., Harvey, D. J., Aplin, R., Shotton, D. M., Dwek, R. A., Barclay, A. N., and Driscoll, P. C. (1998) Structural analysis of the CD5 antigen—expression, disulphide bond analysis and physical characterisation of the CD5 scavenger receptor superfamily domain 1. Eur. J. Biochem. 257, 131–141.

    Article  PubMed  CAS  Google Scholar 

  40. Morgan, W. D., Birdsall, B., Frenkiel, T. A., Gradwell, M. G., Burghaus, P. A., Syed, S. E., Uthaipibull, C., Holder, A. A., and Feeney, J. (1999) Solution structure of an EGF module pair from the Plasmodium falciparum merozoite surface protein 1. J. Mol. Biol. 289, 113–122.

    Article  PubMed  CAS  Google Scholar 

  41. Wiles, A. P., Shaw, G., Bright, J., Perczel, A., Campbell, I. D., and Barlow, P. N. (1997) NMR studies of a viral protein that mimics the regulators of complement activation. J. Mol. Biol. 272, 253–265.

    Article  PubMed  CAS  Google Scholar 

  42. Denton, H., Smith, M., Husi, H., Uhrin, D., Barlow, P. N., Batt, C. A., and Sawyer, L. (1998) Isotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pastoris. Prot. Exp. Purif. 14, 97–103.

    Article  CAS  Google Scholar 

  43. Uhrinova, S., Uhrin, D., Denton, H., Smith, M., Sawyer, L., and Barlow, P. N. (1998) Complete assignment of 1H, 13C and 15N chemical shifts for bovine beta-lactoglobulin: secondary structure and topology of the native state is retained in a partially unfolded form. J. Biomol. NMR 12, 89–107.

    Article  PubMed  CAS  Google Scholar 

  44. Kim, T. R., Goto, Y., Hirota, N., Kuwata, K., Denton, H., Wu, S. Y., Sawyer, L., and Batt, C. A. (1997) High-level expression of bovine beta-lactoglobulin in Pichia pastoris and characterization of its physical properties. Protein Eng. 10, 1339–1345.

    Article  PubMed  CAS  Google Scholar 

  45. Mine, S., Ueda, T., Hashimoto, Y., Tanaka, Y., and Imoto, T. (1999) High-level expression of uniformly 15N-labeled hen lysozyme in Pichia pastoris and identification of the site in hen lysozyme where phosphate ion binds using NMR measurements. FEBS Lett. 448, 33–37.

    Article  PubMed  CAS  Google Scholar 

  46. Wood, M. J. and Komives, E. A. (1999) Production of large quantities of isotopically labeled protein in Pichia pastoris by fermentation. J. Biomol. NMR 13, 149–159.

    Article  PubMed  CAS  Google Scholar 

  47. McAlister, M. S., Davis, B., Pfuhl, M., and Driscoll, P. C. (1998) NMR analysis of the N-terminal SRCR domain of human CD5: engineering of a glycoprotein for superior characteristics in NMR experiments. Protein Eng. 11, 847–853.

    Article  PubMed  CAS  Google Scholar 

  48. Nilsen, S. L., DeFord, M. E., Prorok, M., Chibber, B. A., Bretthauer, R. K., and Castellino, F. J. (1997) High-level secretion in Pichia pastoris and biochemical characterization of the recombinant kringle 2 domain of tissue-type plasminogen activator. Biotechnol. Appl. Biochem. 25, 63–74.

    PubMed  CAS  Google Scholar 

  49. Greenwald, J., Le, V., Corrigan, A., Fischer, W., Komives, E., Vale, W., and Choe, S. (1998) Characterization of the extracellular ligand-binding domain of the type II activin receptor. Biochemistry 37, 16711–16718.

    Article  PubMed  CAS  Google Scholar 

  50. Greenwald, J., Fischer, W. H., Vale, W. W., and Choe, S. (1999) Three-finger toxin fold for the extracellular ligand-binding domain of the type II activin receptor serine kinase. Nat. Struct. Biol. 6, 18–22.

    Article  PubMed  CAS  Google Scholar 

  51. Raymond, C. K., Bukowski, T., Holderman, S. D., Ching, A. F. T., Vanaja, E., and Stamm, M. R. (1997) Development of the methylotrophic yeast, Pichia methanolica, for the expression of the 65-kilodalton isoform of human glutamate decarboxylase. Yeast 14, 11–23.

    Article  Google Scholar 

  52. Raymond, C. K. (1999) Recombinant protein expression in Pichia methanolica. In Gene Expression Systems (Fernandez, J. and Hoeffler, J., ed.), Academic Press, San Diego, CA, pp. 193–209.

    Google Scholar 

  53. Montesino, R., Garcia, R., Quintero, O., and Cremata, J. A. (1998) Variation in N-linked oligosaccharide structures on heterologous proteins secreted by the methylotrophic yeast Pichia pastoris. Protein Expr. Purif. 14, 197–207.

    Article  PubMed  CAS  Google Scholar 

  54. Paifer, E., Margolles, E., Cremata, J., Montesino, R., Herrera, L., and Delgado, J. M. (1994) Efficient expression and secretion of recombinant alpha amylase in Pichia pastoris using two different signal sequences. Yeast 10, 1415–1419.

    Article  PubMed  CAS  Google Scholar 

  55. Despreaux, C. W. and Manning, R. F. (1993) The dacA gene of Bacillus stearothemophilus coding for D-alanine carboxypeptidase: cloning, structure and expression in Escherichia coli and Pichia pastoris. Gene 131, 35–41.

    Article  PubMed  CAS  Google Scholar 

  56. Smith, L. A. (1998) Development of recombinant vaccines for botulinum neurotoxin. Toxicon 36, 1539–1548.

    Article  PubMed  CAS  Google Scholar 

  57. Potter, K. J., Bevins, M. A., Vassilieva, E. V., Chiruvolu, V. R., Smith, T., Smith, L. A., and Meagher, M. M. (1998) Production and purification of the heavy-chain fragment C of botulinum neurotoxin, serotype B, expressed in the methylotrophic yeast Pichia pastoris. Protein Expr. Purif. 13, 357–365.

    Article  PubMed  CAS  Google Scholar 

  58. Byrne, M. P., Smith, T. J., Montgomery, V. A., and Smith, L. A. (1998) Purification, potency, and efficacy of the botulinum neurotoxin type A binding domain from Pichia pastoris as a recombinant vaccine candidate. Infect. Immun. 66, 4817–4822.

    PubMed  CAS  Google Scholar 

  59. Rodriguez, E., Han, Y., and Lei, X. G. (1999) Cloning, sequencing, and expression of an Escherichia coli acid phosphatase/phytase gene (appA2) isolated from pig colon. Biochem. Biophys. Res. Commun. 257, 117–123.

    Article  PubMed  CAS  Google Scholar 

  60. Chan, V. J., Selzer, P. M., McKerrow, J. H., and Sakanari, J. A. (1999) Expression and alteration of the S2 subsite of the Leishmania major cathepsin B-like cysteine protease. Biochem. J. 340, 113–117.

    Article  PubMed  CAS  Google Scholar 

  61. Miele, R. G., Prorok, M., Costa, V. A., and Castellino, F. J. (1999) Glycosylation of asparagine-28 of recombinant staphylokinase with high-mannose-type oligosaccharides results in a protein with highly attenuated plasminogen activator activity. J. Biol. Chem. 274, 7769–7776.

    Article  PubMed  CAS  Google Scholar 

  62. Hagenson, M. J., Holden, K. A., Parker, K. A., Wood, P. J., Cruze, J. A., Fuke, M., Hopkins, T. R., and Stroman, D. W. (1989) Expression of streptokinase in Pichia pastoris yeast. Enzyme Microb. Technol. 11, 650–656.

    Article  CAS  Google Scholar 

  63. Markaryan, A., Beall, C. J., and Kolattukudy, P. E. (1996) Inhibition of Aspergillus serine proteinase by Streptomyces subtilisin inhibitor and high-level expression of this inhibitor in Pichia pastoris. Biochem. Biophys. Res. Commun. 220, 372–376.

    Article  PubMed  CAS  Google Scholar 

  64. Thomas, L. and Crawford, D. L. (1998) Cloning of clustered Streptomyces viridosporus T7A lignocellulose catabolism genes encoding peroxidase and endoglucanase and their extracellular expression in Pichia pastoris. Can. J. Microbiol. 44, 364–372.

    Article  PubMed  CAS  Google Scholar 

  65. Biemans, R., Gregoire, D., Haumont, M., Bosseloir, A., Garcia, L., Jacquet, A., Dubeaux, C., and Bollen, A. (1998) The conformation of purified Toxoplasma gondii SAG1 antigen, secreted from engineered Pichia pastoris, is adequate for serorecognition and cell proliferation. J. Biotechnol. 66, 137–146.

    Article  PubMed  CAS  Google Scholar 

  66. Trucksis, M., Conn, T. L., Fasano, A., and Kaper, J. B. (1997) Production of Vibrio cholerae accessory cholera enterotoxin (Ace) in the yeast Pichia pastoris. Infect. Immun. 65, 4984–4988.

    PubMed  CAS  Google Scholar 

  67. De Vouge, M. W., Thaker, A. J., Curran, I. H., Zhang, L., Muradia, G., Rode, H., and Vijay, H. M. (1996) Isolation and expression of a cDNA clone encoding an Alternaria alternata Alt a 1 subunit. Int. Arch. Allergy Immunol. 111, 385–395.

    Article  PubMed  Google Scholar 

  68. Fierobe, H.-P., Mirgorodskaya, E., Frandsen, T. P., Roepstorff, P., and Svensson, B. (1997) Over-expression and characterization of Aspergillus awamori wild-type and mutant glucoamylase secreted by the methylotrophic yeast Pichia pastoris: comparison with wild-type recombinant glucoamylase produced using Saccharomyces cerevisiae and Aspergillus niger as hosts. Protein Expr. Purif. 9, 159–170.

    Article  PubMed  CAS  Google Scholar 

  69. Heimo, H., Palmu, K., and Suominen, I. (1997) Expression in Pichia pastoris and purification of Aspergillus awamori glucoamylase catalytic domain. Protein Expr. Purif. 11, 304.

    Article  Google Scholar 

  70. Calera, J. A., Paris, S., Monod, M., Hamilton, A. J., Debeaupuis, J. P., Diaquin, M., Lopez-Medrano, R., Leal, F., and Latge, J. P. (1997) Cloning and disruption of the antigenic catalase gene of Aspergillus fumigatus. Infect. Immun. 65, 4718–4724.

    PubMed  CAS  Google Scholar 

  71. Beauvais, A., Monod, M., Wyniger, J., Debeaupuis, J. P., Grouzmann, E., Brakch, N., Svab, J., Hovanessian, A. G., and Latge, J. P. (1997) Dipeptidyl-peptidase IV secreted by Aspergillus fumigatus, a fungus pathogenic to humans. Infect. Immun. 65, 3042–3047.

    PubMed  CAS  Google Scholar 

  72. Beauvais, A., Monod, M., Debeaupuis, J. P., Diaquin, M., Kobayashi, H., and Latge, J. P. (1997) Biochemical and antigenic characterization of a new dipeptidyl-peptidase isolated from Aspergillus fumigatus. J. Biol. Chem. 272, 6238–6244.

    Article  PubMed  CAS  Google Scholar 

  73. Martíncz-Ruiz, A., Martínez del Pozo, A., Lacadena, J., Mancheño, J. M., Oñaderra, M., López-Otin, C., and Gavilanes, J. G. (1998) Secretion of recombinant pro- and mature fungal alpha-sarcin ribotoxin by the methylotrophic yeast Pichia pastoris: the Lys-Arg motif is required for maturation. Protein Expr. Purif. 12, 315–322.

    Article  Google Scholar 

  74. Han, Y. and Lei, X. G. (1999) Role of glycosylation in the functional expression of an Aspergillus niger phytase (phyA) in Pichia pastoris. Arch. Biochem. Biophys. 364, 83–90.

    Article  PubMed  CAS  Google Scholar 

  75. Handumrongkul, C., Ma, D. P., and Silva, J. L. (1998) Cloning and expression of Candida guilliermondii xylose reductase gene (xyll) in Pichia pastoris. Appl. Microbiol. Biotechnol. 49, 399–404.

    Article  PubMed  CAS  Google Scholar 

  76. Brocca, S., Schmidt-Dannert, C., Lotti, M., Alberghina, L., and Schmid, R. D. (1998) Design, total synthesis, and functional overexpression of the Candida rugosa lip1 gene coding for a major industrial lipase. Protein Sci. 7, 1415–1422.

    PubMed  CAS  Google Scholar 

  77. Guo, W., Gonzalez-Candelas, L., and Kolattukudy, P. E. (1995) Cloning of a novel constitutively expressed pectate lyase gene pelB from Fusarium solani f. sp. pisi (Nectria haematococca, mating type VI) and characterization of the gene product expressed in Pichia pastoris. J. Bacteriol. 177, 7070–7077.

    PubMed  CAS  Google Scholar 

  78. Guo, W., Gonzalez-Candelas, L., and Kolattukudy, P. E. (1996) Identification of a novel pelD gene expressed uniquely in planta by Fusarium solani f. sp. pisi (Nectria haematococca, mating type VI) and characterization of its protein product as an endopectate lyase. Arch. Biochem. Biophys. 332, 305–312.

    Article  PubMed  CAS  Google Scholar 

  79. Holmquist, M., Tessier, D. C., and Cygler, M. (1997) High-level production of recombinant Geotrichum candidum lipases in yeast Pichia pastoris. Protein Expr. Purif. 11, 35–40.

    Article  PubMed  CAS  Google Scholar 

  80. O’Donohue, M. J., Boissy, G., Huet, J. C., Nespoulous, C., Brunie, S., and Pernollet, J. C. (1996) Overexpression in Pichia pastoris and crystallization of an elicitor protein secreted by the phytopathogenic fungus, Phytophthora cryptogea. Protein Expr. Purif. 8, 254–261.

    Article  PubMed  CAS  Google Scholar 

  81. Minning, S., Schmidt-Dannert, C., and Schmid, R. D. (1998) Functional expression of Rhizopus oryzae lipase in Pichia pastoris: high-level production and some properties. J. Biotechnol. 66, 147–156.

    Article  PubMed  CAS  Google Scholar 

  82. Romero, P. A., Lussier, M., Sdicu, A. M., Bussey, H., and Herscovics, A. (1997) Ktr1p is an alpha-1,2-mannosyltransferase of Saccharomyces cerevisiae. Comparison of the enzymic properties of soluble recombinant Ktr1p and Kre2p/Mnt1p produced in Pichia pastoris. Biochem. J. 321, 289–295.

    PubMed  CAS  Google Scholar 

  83. Chen, H. and McCormick, D. B. (1997) Riboflavin 5′-hydroxymethyl oxidation. Molecular cloning, expression, and glycoprotein nature of the 5′-alde-hyde-forming enzyme from Schizophyllum commune. J. Biol. Chem. 272, 20077–20081.

    Article  PubMed  CAS  Google Scholar 

  84. Jonsson, L. J., Saloheimo, M., and Penttila, M. (1997) Laccase from the white-rot fungus Trametes versicolor: cDNA cloning of lcc1 and expression in Pichia pastoris. Curr. Genet. 32, 425–430.

    Article  PubMed  CAS  Google Scholar 

  85. Bom, I. J., Dielbandhoesing, S. K., Harvey, K. N., Oomes, S. J., Klis, F. M., and Brul, S. (1998) A new tool for studying the molecular architecture of the fungal cell wall: one-step purification of recombinant trichoderma beta-(1-6)-glucanase expressed in Pichia pastoris. Biochim. Biophys. Acta 1425, 419–424.

    PubMed  CAS  Google Scholar 

  86. Hansen, O. C. and Stougaard, P. (1997) Hexose oxidase from the red alga Chondrus crispus. Purification, molecular cloning, and expression in Pichia pastoris. J. Biol. Chem. 272, 11581–11587.

    Article  PubMed  CAS  Google Scholar 

  87. Bojsen, K., Yu, S., Kragh, K. M., and Marcussen, J. (1999) A group of alpha-1,4-glucan lyases and their genes from the red alga Gracilariopsis lemaneiformis: purification, cloning, and heterologous expression. Biochim. Biophys. Acta 1430, 396–402.

    PubMed  CAS  Google Scholar 

  88. Kocken, C. H. M., Dubbeld, M. A., Van Der Wel, A., Pronk, J. T., Waters, A. P., Langermans, J. A. M., and Thomas, A. W. (1999) High-level expression of Plasmodium vivax apical membrane antigen 1 (AMA-1) in Pichia pastoris: strong immunogenicity in Macaca mulatta immunized with P. vivax AMA-1 and adjuvant SBAS2. Infect. Immun. 67, 43–49.

    PubMed  CAS  Google Scholar 

  89. Linder, S., Schliwa, M., and Kube-Granderath, E. (1997) Expression of Reticulomyxa filosa tubulins in Pichia pastoris: regulation of tubulin pools. FEBS Lett. 417, 33–37.

    Article  PubMed  CAS  Google Scholar 

  90. Vandersall-Nairn, A. S., Merkle, R. K., O’Brien, K., Oeltmann, T. N., and Moremen, K. W. (1998) Cloning, expression, purification, and characterization of the acid alpha-mannosidase from Trypanosoma cruzi. Glycobiology 8, 1183–1194.

    Article  PubMed  CAS  Google Scholar 

  91. Weik, R., Francky, A., Striedner, G., Raspor, P., Bayer, K., and Mattanovich, D. (1998) Recombinant expression of alliin lyase from garlic (Allium sativum) in bacteria and yeasts. Planta Med. 64, 387–388.

    Article  PubMed  CAS  Google Scholar 

  92. Su, W., Mertens, J. A., Kanamaru, K., Campbell, W. H., and Crawford, N. M. (1997) Analysis of wild-type and mutant plant nitrate reductase expressed in the methylotrophic yeast Pichia pastoris. Plant Physiol. 115, 1135–1143.

    Article  PubMed  CAS  Google Scholar 

  93. Su, W., Huber, S. C., and Crawford, N. M. (1996) Identification in vitro of a post-translational regulatory site in the hinge 1 region of Arabidopsis nitrate reductase. Plant Cell 8, 519–527.

    Article  PubMed  CAS  Google Scholar 

  94. Hochstrasser, U., Luscher, M., De Virgilio, C., Boler, T., and Wiemken, A. (1998) Expression of a functional barley sucrose-fructan 6-fructosyltransferase in the methylotrophic yeast Pichia pastoris. FEBS Lett. 440, 356–360.

    Article  PubMed  CAS  Google Scholar 

  95. Juge, N., Andersen, J. S., Tull, D., Roepstorff, P., and Svensson, B. (1996) Overexpression, purification, and characterization of recombinant barley alpha-amylases 1 and 2 secreted by the methylotrophic yeast Pichia pastoris. Protein Expr. Purif. 8, 204–214.

    Article  PubMed  CAS  Google Scholar 

  96. Tibbot, B. K., Henson, C. A., and Skadsen, R. W. (1998) Expression of enzymatically active, recombinant barley alpha-glucosidase in yeast and immunological detection of alpha-glucosidase from seed tissue. Plant Mol. Biol. 38, 379–391.

    Article  PubMed  CAS  Google Scholar 

  97. Zhu, A., Monahan, C., Zhang, Z., Hurst, R., Leng, L., and Goldstein, J. (1995) High-level expression and purification of coffee bean alpha-galactosidase produced in the yeast Pichia pastoris. Arch. Biochem. Biophys. 324, 65–70.

    Article  PubMed  CAS  Google Scholar 

  98. White, P. C., Cordeiro, M. C., Arnold, D., Brodelius, P. E., and Kay, J. (1999) Processing, activity, and inhibition of recombinant cyprosin, an aspartic proteinase from cardoon (Cynara cardunculus). J. Biol. Chem. 274, 16685–16693.

    Article  PubMed  CAS  Google Scholar 

  99. Chang, Z. N., Peng, H. J., Lee, W. C., Chen, T. S., Chua, K. Y., Tsai, L. C., Chi, C. W., and Han, S. H. (1999) Sequence polymorphism of the group 1 allergen of Bermuda grass pollen. Clin. Exp. Allergy 29, 488–496.

    Article  PubMed  CAS  Google Scholar 

  100. Smith, P. M., Suphioglu, C., Griffith, I. J., Theriault, K., Knox, R. B., and Singh, M. B. (1996) Cloning and expression in yeast Pichia pastoris of a biologically active form of Cyn d 1, the major allergen of Bermuda grass pollen. J. Allergy Clin. Immunol. 98, 331–343.

    Article  PubMed  CAS  Google Scholar 

  101. Raemaekers, R. J. M., de Muro, L., Gatehouse, J. A., and Fordham-Skelton, A. P. (1999) Functional phytohaemagglutinin (PHA) and Galanthus nivalis agglutinin (GNA) expressed in Pichia pastoris: correct N-terminal processing and secretion of heterologous proteins expressed using the PHA-E signal peptide. Eur. J. Biochem. 265, 394–403.

    Article  PubMed  CAS  Google Scholar 

  102. Hasslacher, M., Schall, M., Hayn, M., Bona, R., Rumbold, K., Luckl, J., Griengl, H., Kohlwein, S. D., and Schwab, H. (1997) High-level intracellular expression of hydroxynitrile lyase from the tropical rubber tree Hevea brasiliensis in microbial hosts. Protein Expr. Purif. 11, 61–71.

    Article  PubMed  CAS  Google Scholar 

  103. Sowka, S., Wagner, S., Krebitz, M., Arija-Mad-Arif, S., Yusof, F., Kinaciyan, T., Brehler, R., Scheiner, O., and Breiteneder, H. (1998) cDNA cloning of the 43-kDa latex allergen Hev b7 with sequence similarity to patatins and its expression in the yeast Pichia pastoris. Eur. J. Biochem. 255, 213–219.

    Article  PubMed  CAS  Google Scholar 

  104. Breiteneder, H., Sowka, S., Wagner, S., Krebitz, M., Hafner, C., Kinaciyan, T., Yeang, H. Y., and Scheiner, O. (1999) Cloning of the patatin-like latex allergen Hev b 7, its expression in the yeast Pichia pastoris and its immunological characterization. Int. Arch. Allergy Immunol. 118, 309–310.

    Article  PubMed  CAS  Google Scholar 

  105. Morris, R. O., Bilyeu, K. D., Laskey, J. G., and Cheikh, N. N. (1999) Isolation of a gene encoding a glycosylated cytokinin oxidase from maize. Biochem. Biophys. Res. Commun. 255, 328–333.

    Article  PubMed  CAS  Google Scholar 

  106. Mozley, D., Remberg, A., and Gartner, W. (1997) Large-scale generation of affinity-purified recombinant phytochrome chromopeptide. Photochem. Photobiol. 66, 710–715.

    PubMed  CAS  Google Scholar 

  107. Kneip, C., Mozley, D., Hildebrandt, M. P., Gartner, W., Braslavsky, S. E., and Schaffner, K. (1997) Effect of chromophore exchange on the resonance Raman spectra of recombinant phytochromes. FEBS Lett. 414, 23–26.

    Article  PubMed  CAS  Google Scholar 

  108. Remberg, A., Ruddat, A., Braslavsky, S. E., Gartner, W., and Schaffner, K. (1998) Chromophore incorporation, Pr to Pfr kinetics, and Pfr thermal reversion of recombinant N-terminal fragments of phytochrome A and B chromoproteins. Biochemistry 37, 9983–9990.

    Article  PubMed  CAS  Google Scholar 

  109. Huecas, S., Villalba, M., Gonzalez, E., Martinez-Ruiz, A., and Rodriguez, R. (1999) Production and detailed characterization of biologically active olive pollen allergen Ole e 1 secreted by the yeast Pichia pastoris. Eur. J. Biochem. 261, 539–546.

    Article  PubMed  CAS  Google Scholar 

  110. Ferrarese, L., Trainotti, L., Gattolin, S., and Casadoro, G. (1998) Secretion, purification and activity of two recombinant pepper endo-beta-1,4-glucanases expressed in the yeast Pichia pastoris. FEBS Lett. 422, 23–26.

    Article  PubMed  CAS  Google Scholar 

  111. Sowka, S., Hsieh, L. S., Krebitz, M., Akasawa, A., Martin, B. M., Starrett, D., Peterbauer, C. K., Scheiner, O., and Breiteneder, H. (1998) Identification and cloning of prs a 1, a 32-kDa endochitinase and major allergen of avocado, and its expression in the yeast Pichia pastoris. J. Biol. Chem. 273, 28091–28097.

    Article  PubMed  CAS  Google Scholar 

  112. Grobe, K., Becker, W.-M., Schlaak, M., Petersen, A. (1999) Grass group I allergens (beta-expansins) are novel, papain-related proteinases. Eur. J. Biochem. 263, 33–40.

    Article  PubMed  CAS  Google Scholar 

  113. Ruddat, A., Schmidt, P., Gatz, C., Braslavsky, S. E., Gartner, W., and Schaffner, K. (1997) Recombinant type A and B phytochromes from potato. Transient absorption spectroscopy. Biochemistry 36, 103–111.

    Article  PubMed  CAS  Google Scholar 

  114. Hiller, K. M., Lubahn, B. C., and Klapper, D. G. (1998) Cloning and expression of ragweed allergen Amb a 6. Scand. J. Immunol. 48, 26–36.

    Article  PubMed  CAS  Google Scholar 

  115. Penheiter, A. R., Klucas, R. V., and Sarath, G. (1998) Purification and characterization of a soybean root nodule phosphatase expressed in Pichia pastoris. Protein Expr. Purif. 14, 125–130.

    Article  PubMed  CAS  Google Scholar 

  116. Payne, M. S., Petrillo, K. L., Gavagan, J. E., Wagner, L. W., DiCosimo, R., and Anton, D. L. (1995) High-level production of spinach glycolate oxidase in the methylotrophic yeast Pichia pastoris: engineering a biocatalyst. Gene 167, 215–219.

    Article  PubMed  CAS  Google Scholar 

  117. Payne, M. S., Petrillo, K. L., Gavagan, J. E., DiCosimo, R., Wagner, L. W., and Anton, D. L. (1997) Engineering Pichia pastoris for biocatalysis. co-production of two active enzymes. Gene 194, 179–182.

    Article  PubMed  CAS  Google Scholar 

  118. Brandes, H. K., Hartman, F. C., Lu, T. Y., and Larimer, F. W. (1996) Efficient expression of the gene for spinach phosphoribulokinase in Pichia pastoris and utilization of the recombinant enzyme to explore the role of regulatory cysteinyl residues by site-directed mutagenesis. J. Biol. Chem. 271, 6490–6496.

    Article  PubMed  CAS  Google Scholar 

  119. Kristensen, A. K., Brunstedt, J., Nielsen, J. E., Mikkelsen, J. D., Roepstorff, P., and Nielsen, K. K. (1999) Processing, disulfide pattern, and biological activity of a sugar beet defensin, AX2, expressed in Pichia pastoris. Protein Expr. Purif. 16, 377–387.

    Article  PubMed  CAS  Google Scholar 

  120. Petersen, A., Grobe, K., Lindner, B., Schlaak, M., and Becker, W. M. (1997) Comparison of natural and recombinant isoforms of grass pollen allergens. Electrophoresis 18, 819–825.

    Article  PubMed  CAS  Google Scholar 

  121. Brenner, E. D., Lambert, K. N., Kaloshian, I., and Williamson, V. M. (1998) Characterization of LeMir, a root-knot nematode-induced gene in tomato with an encoded product secreted from the root. Plant Physiol. 118, 237–247.

    Article  PubMed  CAS  Google Scholar 

  122. Klein, C., de Lamotte-Guery, F., Gautier, F., Moulin, G., Boze, H., Joudrier, P., and Gautier, M. F. (1998) High-level secretion of a wheat lipid transfer protein in Pichia pastoris. Protein Expr. Purif. 13, 73–82.

    Article  PubMed  CAS  Google Scholar 

  123. Ogawa, M., Nakamura, S., Atsuchi, T., Tamiya, T., Tsuchiya, T., and Nakai, S. (1999) Macromolecular antimicrobial glycoprotein, achacin, expressed in a methylotrophic yeast Pichia pastoris. FEBS Lett. 448, 41–44.

    Article  PubMed  CAS  Google Scholar 

  124. Munshi, C. and Lee, H. C. (1997) High-level expression of recombinant Aplysia ADP-ribosyl cyclase in Pichia pastoris by fermentation. Protein Expr. Purif. 11, 104–110.

    Article  PubMed  CAS  Google Scholar 

  125. Monosov, E. Z., Wenzel, T. J., Luers, G. H., Heyman, J. A., and Subramani, S. (1996) Labeling of peroxisomes with green fluorescent protein in living P. pastoris cells. J. Histochem. Cytochem. 44, 581–589.

    PubMed  CAS  Google Scholar 

  126. Gallet, P. F., Vaujour, H., Petit, J. M., Maftah, A., Oulmouden, A., Oriol, R., Le Narvor, C., Guilloton, M., and Julien, R. (1998) Heterologous expression of an engineered truncated form of human Lewis fucosyltransferase (Fuc-TIII) by the methylotrophic yeast Pichia pastoris. Glycobiology 8, 919–925.

    Article  PubMed  CAS  Google Scholar 

  127. Garcia-Garcia, J. C., Soto, A., Nigro, F., Mazza, M., Joglar, M., Hechevarria, M., Lamberti, J., and de la Fuente, J. (1998) Adjuvant and immunostimulating properties of the recombinant Bm86 protein expressed in Pichia pastoris. Vaccine 16, 1053–1055.

    Article  PubMed  CAS  Google Scholar 

  128. Garcia-Garcia, J. C., Montero, C., Rodriquez, M., Soto, A., Redondo, M., Valdes, M., Mendez, L., and de la Fuente, J. (1998) Effect of particulation on the immunogenic and protective properties of the recombinant Bm86 antigen expressed in Pichia pastoris. Vaccine 16, 374–380.

    Article  PubMed  CAS  Google Scholar 

  129. Rodríguez, M., Rubiera, R., Penichet, M., Montesinos, R., Cremata, J., Falcón, V., Sánchez, G., Bringas, R., Cordovés, C., Valdés, M., Lleonart, R., Herrera, L., and de la Fuente, J. (1994) High level expression of the B. microplus Bm86 antigen in the yeast Pichia pastoris forming highly immunogenic particles for cattle. J. Biotechnol. 33, 135–146.

    Article  PubMed  Google Scholar 

  130. Vailes, L. D., Kinter, M. T., Arruda, L. K., and Chapman, M. D. (1998) High-level expression of cockroach allergen, Bla g 4, in Pichia pastoris. J. Allergy Clin. Immunol. 101, 274–280.

    Article  CAS  Google Scholar 

  131. Williams, T. A., Michaud, A., Houard, X., Chauvet, M. T., Soubrier, F., and Corvol, P. (1996) Drosophila melanogaster angiotensin I-converting enzyme expressed in Pichia pastoris resembles the C domain of the mammalian homologue and does not require glycosylation for secretion and enzymic activity. Biochem. J. 318, 125–131.

    PubMed  CAS  Google Scholar 

  132. Jackson, V. N., Cameron, J. M., Zammit, V. A., and Price, N. T. (1999) Sequencing and functional expression of the malonyl-CoA-sensitive carnitine palmitoyltransferase from Drosophila melanogaster. Biochem. J. 341, 483–489.

    Article  PubMed  CAS  Google Scholar 

  133. McCollum, D., Monosov, E., and Subramani, S. (1993) The pas8 mutant of Pichia pastoris exhibits the peroxisomal protein import deficiencies of Zellweger syndrome—the PAS8 protein binds to the COOH-terminal tripeptide peroxisomal targeting signal, and is a member of the TPR protein family. J. Cell Biol. 121, 761–774.

    Article  PubMed  CAS  Google Scholar 

  134. Canales, M., Enriquez, A., Ramos, E., Cabrera, D., Dandie, H., Soto, A., Falcon, V., Rodriguez, M., and de la Fuente, J. (1997) Large-scale production in Pichia pastoris of the recombinant vaccine Gavac against cattle tick. Vaccine 15, 414–422.

    Article  PubMed  CAS  Google Scholar 

  135. Brankamp, R. G., Sreekrishna, K., Smith, P. L., Blankenship, D. T., and Cardin, A. D. (1995) Expression of a synthetic gene encoding the anticoagulant-antimetastatic protein ghilanten by the methylotropic yeast Pichia pastoris. Protein Expr. Purif. 6, 813–820.

    Article  PubMed  CAS  Google Scholar 

  136. Rosenfeld, S. A., Nadeau, D., Tirado, J., Hollis, G. F., Knabb, R. M., and Jia, S. (1996) Production and purification of recombinant hirudin expressed in the methylotrophic yeast Pichia pastoris. Protein Expr. Purif. 8, 476–482.

    Article  PubMed  CAS  Google Scholar 

  137. Briand, L., Perez, V., Huet, J. C., Danty, E., Masson, C., and Pernollet, J. C. (1999) Optimization of the production of a honeybee odorant-binding protein by Pichia pastoris. Protein Expr. Purif. 15, 362–369.

    Article  PubMed  CAS  Google Scholar 

  138. Danty, E., Briand, L., Michard-Vanhee, C., Perez, V., Arnold, G., Gaudemer, O., Huet, D., Huet, J.-C., Ouali, C., Masson, C., and Pernollet, J.-C. (1999) Cloning and expression of a queen pheromone-binding protein in the honeybee: an olfactory-specific, developmentally regulated protein. J. Neuroscience 19, 7468–7475.

    CAS  Google Scholar 

  139. Hussein, A. S., Chacon, M. R., Smith, A. M., Tosado-Acevedo, R., and Selkirk, M. E. (1999) Cloning, expression, and properties of a nonneuronal secreted acetylcholinesterase from the parasitic nematode Nippostrongylus brasiliensis. J. Biol. Chem. 274, 9312–9319.

    Article  PubMed  CAS  Google Scholar 

  140. Fahnestock, S. R. and Bedzyk, L. A. (1997) Production of synthetic spider dragline silk protein in Pichia pastoris. Appl. Microbiol. Biotechnol. 47, 33–39.

    Article  PubMed  CAS  Google Scholar 

  141. Monsalve, R. I., Lu, G., and King, T. P. (1999) Expressions of recombinant venom allergen, antigen 5 of yellowjacket (Vespula vulgaris) and paper wasp (Polistes annularis), in bacteria or yeast. Protein Expr. Purif. 16, 410–416.

    Article  PubMed  CAS  Google Scholar 

  142. Vozza, L. A., Wittwer, L., Higgins, D. R., Purcell, T. J., Bergseid, M., Collins-Racie, L. A., LaVallie, E. R., and Hoeffler, J. P. (1996) Production of a recombinant bovine enterokinase catalytic subunit in the methylotrophic yeast Pichia pastoris. Biotechnology (NY) 14, 77–81.

    Article  CAS  Google Scholar 

  143. Samaddar, M., Catterall, J. F., and Dighe, R. R. (1997) Expression of biologically active beta subunit of bovine follicle-stimulating hormone in the methylotrophic yeast Pichia pastoris. Protein Expr. Purif. 10, 345–355.

    Article  PubMed  CAS  Google Scholar 

  144. Rodriguez, M., Martinez, V., Alazo, K., Suarez, M., Redondo, M., Montero, C., Besada, V., and de la Fuente, J. (1998) The bovine IFN-omega 1 is biologically active and secreted at high levels in the yeast Pichia pastoris. J. Biotechnol. 60, 3–14.

    Article  PubMed  CAS  Google Scholar 

  145. Digan, M. E., Lair, S. V., Brierley, R. A., Siegel, R. S., Williams, M. E., Ellis, S. B., Kellaris, P. A., Provow, S. A., Craig, W. S., Veliçelebi G., Harpold, M., and Thill, G. P. (1989) Continuous production of a novel lysozyme via secretion from the yeast, Pichia pastoris. Biotechnology (NY) 7, 160–164.

    Article  CAS  Google Scholar 

  146. Abdulaev, N. G., Popp, M. P., Smith, W. C., and Ridge, K. D. (1997) Functional expression of bovine opsin in the methylotrophic yeast Pichia pastoris. Protein Expr. Purif. 10, 61–69.

    Article  PubMed  CAS  Google Scholar 

  147. Vedvick, T., Buckholtz, R. G., Engel, M., Urcan, M., Kinney, J., Provow, S., Siegel, R. S., and Thill, G. P. (1991) High-level secretion of biologically active aprotinin from the yeast Pichia pastoris. J. Ind. Microbiol. 7, 197–201.

    Article  PubMed  CAS  Google Scholar 

  148. Choi, B.-K. and Jimenez-Flores, R. (1996) Study of putative glycosylation site in bovine beta-casein introduced by PCR-based site-directed mutagenesis. J. Agric. Food Chem. 44, 358–364.

    Article  CAS  Google Scholar 

  149. Johnsen, L. B., Ravn, P., Berglund, L., Petersen, T. E., Rasmussen, L. K., Heegaard, C. W., Rasmussen, J. T., Benfeldt, C., and Fedosov, S. N. (1998) A refined kinetic analysis of plasminogen activation by recombinant bovine tissue-type plasminogen activator indicates two interconvertible activator forms. Biochemistry 37, 12631–12639.

    Article  PubMed  CAS  Google Scholar 

  150. Fedosov, S. N., Berglund, L., Nexo, E., and Petersen, T. E. (1999) Sequence, S-S bridges, and spectra of bovine transcobalamin expressed in Pichia pastoris. J. Biol. Chem. 274, 26015–26020.

    Article  PubMed  CAS  Google Scholar 

  151. Wedlock, D. N., Goh, L. P., McCarthy, A. R., Midwinter, R. G., Parlane, N. A., and Buddle, B. M. (1999) Physiological effects and adjuvanticity of recombinant brushtail possum TNF-alpha. Immunol. Cell Biol. 77, 28–33.

    Article  PubMed  CAS  Google Scholar 

  152. Morel, N. and Massoulie, J. (1997) Expression and processing of vertebrate acetylcholinesterase in the yeast Pichia pastoris. Biochem. J. 328, 121–129.

    PubMed  CAS  Google Scholar 

  153. Zhu, A., Monahan, C., Wang, Z. K., and Goldstein, J. (1996) Expression, purification, and characterization of recombinant alpha-N-acetylgalactosaminidase produced in the yeast Pichia pastoris. Protein Expr. Purif. 8, 456–462.

    Article  PubMed  CAS  Google Scholar 

  154. Simon, S. and Massoulie, J. (1997) Cloning and expression of acetylcholinesterase from Electrophorus. Splicing pattern of the 3′ exons in vivo and in transfected mammalian cells. J. Biol. Chem. 272, 33045–33055.

    Article  PubMed  CAS  Google Scholar 

  155. Rautiainen, J., Auriola, S., Rouvinen, J., Kauppinen, J., Zeiler, T., Novikov, D., Virtanen, T., and Mantyjarvi, R. A. (1998) Molecular and crystal properties of Bos d 2, an allergenic protein of the lipocalin family. Biochem. Biophys. Res. Commun. 247, 746–750.

    Article  PubMed  CAS  Google Scholar 

  156. Weiss, H. M., Haase, W., Michel, H., and Reilander, H. (1998) Comparative biochemical and pharmacological characterization of the mouse 5HT5A 5-hydroxytryptamine receptor and the human beta 2-adrenergic receptor produced in the methylotrophic yeast Pichia pastoris. Biochem. J. 330, 1137–1147.

    PubMed  CAS  Google Scholar 

  157. Werten, M. W., van den Bosch, T. J., Wind, R. D., Mooibroek, H., and de Wolf, F. A. (1999) High-yield secretion of recombinant gelatins by Pichia pastoris. Yeast 15, 1087–1096.

    Article  PubMed  CAS  Google Scholar 

  158. Masure, S., Paemen, L., Van Aelst, I., Fiten, P., Proost, P., Billiau, A., Van Damme, J., and Opdenakker, G. (1997) Production and characterization of recombinant active mouse gelatinase B from eukaryotic cells and in vivo effects after intravenous administration. Eur. J. Biochem. 244, 21–30.

    Article  PubMed  CAS  Google Scholar 

  159. Merkle, R. K., Zhang, Y., Ruest, P. J., Lal, A., Liao, Y. F., and Moremen, K. W. (1997) Cloning, expression, purification, and characterization of the murine lysosomal acid alpha-mannosidase. Biochim. Biophys. Acta 1336, 132–146.

    PubMed  CAS  Google Scholar 

  160. Ferrari, E., Lodi, T., Sorbi, R. T., Tirindelli, R., Cavaggioni, A., and Spisni, A. (1997) Expression of a lipocalin in Pichia pastoris: secretion, purification and binding activity of a recombinant mouse major urinary protein. FEBS Lett. 401, 73–77.

    Article  PubMed  CAS  Google Scholar 

  161. Urbatsch, I. L., Beaudet, L., Carrier, I., and Gros, P. (1998) Mutations in either nucleotide-binding site of P-glycoprotein (Mdr3) prevent vanadate trapping of nucleotide at both sites. Biochemistry 37, 4592–4602.

    Article  PubMed  CAS  Google Scholar 

  162. Beaudet, L., Urbatsch, I. L., and Gros, P. (1998) High-level expression of mouse Mdr3 P-glycoprotein in yeast Pichia pastoris and characterization of ATPase activity. Methods Enzymol. 292, 397–413.

    PubMed  CAS  Google Scholar 

  163. Beaudet, L., Urbatsch, I. L., and Gros, P. (1998) Mutations in the nucleotide-binding sites of P-glycoprotein that affect substrate specificity modulate substrate-induced adenosine triphosphatase activity. Biochemistry 37, 9073–9082.

    Article  PubMed  CAS  Google Scholar 

  164. Eldin, P., Pauza, M. E., Hieda, Y., Lin, G., Murtaugh, M. P., Pentel, P. R., and Pennell, C. A. (1997) High-level secretion of two antibody single chain Fv fragments by Pichia pastoris. J. Immunol. Methods 201, 67–75.

    Article  PubMed  CAS  Google Scholar 

  165. Boehm, T., Pirie-Shepard, S., Trinh, L. B., Shiloach, J., and Folkman, J. (1999) Disruption of the KEX1 gene in Pichia pastoris allows expression of full-length murine and human endostatin. Yeast 15, 563–567.

    Article  PubMed  CAS  Google Scholar 

  166. Vallee, F., Lal, A., Moremen, K. W., and Howell, P. L. (1999) Purification, crystallization and preliminary X-ray crystallographic analysis of recombinant murine Golgi mannosidase IA, a class I alphamannosidase involved in Asn-linked oligosaccharide maturation. Acta Crystallogr. D Biol. Crystallogr. 55, 571–573.

    Article  PubMed  CAS  Google Scholar 

  167. Jerva, L. F., Sullivan, G., and Lolis, E. (1997) Functional and receptor binding characterization of recombinant murine macrophage inflammatory protein 2: sequence analysis and mutagenesis identify receptor binding epitopes. Protein Sci. 6, 1643–1652.

    PubMed  CAS  Google Scholar 

  168. Fidler, A. E., Lun, S., Young, W., and McNatty, K. P. (1998) Expression and secretion of a biologically active glycoprotein hormone, ovine follicle stimulating hormone, by Pichia pastoris. J. Mol. Endocrinol. 21, 327–336.

    Article  PubMed  CAS  Google Scholar 

  169. Ventura, S., Villegas, V., Sterner, J., Larson, J., Vendrell, J., Hershberger, C. L., Aviles, F. X. (1999) Mapping the pro-region of carboxypeptidase B by protein engineering. Cloning, overexpression, and mutagenesis of the porcine proenzyme. J. Biol. Chem. 274, 19925–19933.

    Article  PubMed  CAS  Google Scholar 

  170. Richard, F., Robert, P., Remy, J. J., Martinat, N., Bidart, J. M., Salesse, R., and Combarnous, Y. (1998) High-level secretion of biologically active recombinant porcine follicle-stimulating hormone by the methylotrophic yeast Pichia pastoris. Biochem. Biophys. Res. Commun. 245, 847–852.

    Article  PubMed  CAS  Google Scholar 

  171. Wuebbens, M. W., Roush, E. D., Decastro, C. M., and Fierke, C. A. (1997) Cloning, sequencing, and recombinant expression of the porcine inhibitor of carbonic anhydrase: a novel member of the transferrin family. Biochemistry 36, 4327–4336.

    Article  PubMed  CAS  Google Scholar 

  172. Reddy, R. G., Yoshimoto, T., Yamamoto, S., and Marnett, L. J. (1994) Expression, purification, and characterization of porcine leukocyte 12-lipoxygenase produced in the methylotrophic yeast, Pichia pastoris. Biochem. Biophys. Res. Commun. 205, 381–388.

    Article  PubMed  CAS  Google Scholar 

  173. Doring, F., Theis, S., and Daniel, H. (1997) Expression and functional characterization of the mammalian intestinal peptide transporter PEPT1 in the methylotrophic yeast Pichia pastoris. Biochem. Biophys. Res. Commun. 232, 656–662.

    Article  PubMed  CAS  Google Scholar 

  174. Doring, F., Michel, T., Rosel, A., Nickolaus, M., and Daniel, H. (1998) Expression of the mammalian renal peptide transporter PEPT2 in the yeast Pichia pastoris and applications of the yeast system for functional analysis. Mol. Membr. Biol. 15, 79–88.

    PubMed  CAS  Google Scholar 

  175. Ridder, R., Schmitz, R., Legay, F., and Gram, H. (1995) Generation of rabbit monoclonal antibody fragments from a combinatorial phage display library and their production in the yeast Pichia pastoris. Biotechnology (NY) 13, 255–260.

    Article  CAS  Google Scholar 

  176. Kotake, H., Li, Q., Ohnishi, T., Ko, K. W., Agellon, L. B., and Yokoyama, S. (1996) Expression and secretion of rabbit plasma cholesteryl ester transfer protein by Pichia pastoris. J. Lipid Res. 37, 599–605.

    PubMed  CAS  Google Scholar 

  177. Sadhukhan, R., Sen, G. C., and Sen, I. (1996) Synthesis and cleavage—secretion of enzymatically active rabbit angiotensin-converting enzyme in Pichia pastoris. J. Biol. Chem. 271, 18310–18313.

    Article  PubMed  CAS  Google Scholar 

  178. Heim, J., Schmidt-Dannert, C., Atomi, H., and Schmid, R. D. (1998) Functional expression of a mammalian acetylcholinesterase in Pichia pastoris: comparison to acetylcholinesterase, expressed and reconstituted from Escherichia coli. Biochim. Biophys. Acta 1396, 306–319.

    PubMed  CAS  Google Scholar 

  179. He, C., Alexander, J. J., Lim, A., and Quigg, R. J. (1997) Production of the rat complement regulator, Crry, as an active soluble protein in Pichia pastoris. Arch. Biochem. Biophys. 341, 347–352.

    Article  PubMed  CAS  Google Scholar 

  180. Chen, Y. J. and Gonatas, N. K. (1997) The Golgi sialoglycoprotein MG160, expressed in Pichia pastoris, does not require complex carbohydrates and sialic acid for secretion and basic fibroblast growth factor binding. Biochem. Biophys. Res. Commun. 234, 68–72.

    Article  PubMed  CAS  Google Scholar 

  181. Mistry, A. R., Falciola, L., Monaco, L., Tagliabue, R., Acerbis, G., Knight, A., Harbottle, R. P., Soria, M., Bianchi, M. E., Coutelle, C., and Hart, S. L. (1997) Recombinant HMG1 protein produced in Pichia pastoris: a nonviral gene delivery agent. Biotechniques 22, 718–729.

    PubMed  CAS  Google Scholar 

  182. Zhu, H., Shi, J., de Vries, Y., Arvidson, D. N., Cregg, J. M., and Woldegiorgis, G. (1997) Functional studies of yeast-expressed human heart muscle carnitine palmitoyltransferase I. Arch. Biochem. Biophys. 347, 53–61.

    Article  PubMed  CAS  Google Scholar 

  183. de Vries, Y., Arvidson, D. N., Waterham, H. R., Cregg, J. M., and Woldegiorgis, G. (1997) Functional characterization of mitochondrial carnitine palmitoyltransferases I and II expressed in the yeast Pichia pastoris. Biochemistry 36, 5285–5292.

    Article  PubMed  Google Scholar 

  184. Kasper, C, Rasmussen, H., Berezin, V., Bock, E., and Larsen, I. K. (1999) Expression, crystallization and preliminary X-ray analysis of the two amino-terminal Ig domains of the neural cell adhesion molecule (NCAM). Acta Crystallogr. D Biol. Crystallogr. 55, 1598–1600.

    Article  PubMed  CAS  Google Scholar 

  185. Gachhui, R., Presta, A., Bentley, D. F., Abu-Soud, H. M., McArthur, R., Brudvig, G., Ghosh, D. K., and Stuehr, D. J. (1996) Characterization of the reductase domain of rat neuronal nitric oxide synthase generated in the methylotrophic yeast Pichia pastoris. Calmodulin response is complete within the reductase domain itself. J. Biol. Chem. 271, 20594–20602.

    Article  PubMed  CAS  Google Scholar 

  186. Qin, Y. M., Poutanen, M. H., Helander, H. M., Kvist, A. P., Siivari, K. M., Schmitz, W., Conzelmann, E., Hellman, U., and Hiltunen, J. K. (1997) Peroxisomal multifunctional enzyme of beta-oxidation metabolizing D-3-hydroxyacyl-CoA esters in rat liver: molecular cloning, expression and characterization. Biochem. J. 321, 21–28.

    PubMed  CAS  Google Scholar 

  187. Yu, Y., Vranken, W., Goudreau, N., de Miguel, E., Magny, M. C., Mort, J. S., Dupras, R., Storer, A. C., and Ni, F. (1998) An NMR-based identification of peptide fragments mimicking the interactions of the cathepsin B propeptide. FEBS Lett. 429, 9–16.

    Article  PubMed  CAS  Google Scholar 

  188. Sivaraman, J., Coulombe, R., and Cygler, M. (1996) Crystallization of rat procathepsin B. Acta Crystallogr. 52, 874.

    CAS  Google Scholar 

  189. Loewen, M. C., Liu, X., Davies, P. L., and Daugulis, A. J. (1997) Biosynthetic production of type II fish antifreeze protein: fermentation by Pichia pastoris. Appl. Microbiol. Biotechnol. 48, 480–486.

    Article  PubMed  CAS  Google Scholar 

  190. Trant, J. M. (1996) Functional expression of recombinant spiny dogfish shark (Squalus acanthias) cytochrome P450c17 (17 alpha-hydroxylase/C17,20-lyase) in yeast (Pichia pastoris). Arch. Biochem. Biophys. 326, 8–14.

    Article  PubMed  CAS  Google Scholar 

  191. Weiss, S., Famulok, M., Edenhofer, F., Wang, Y.-H., Jones, I. M., Groschup, M., and Winnacker, E. L. (1995) Overexpression of active Syrian golden hamster prion protein PrPc as a glutathione S-transferase fusion in heterologous systems. J. Virol. 69, 4776–4783.

    PubMed  CAS  Google Scholar 

  192. Tremblay, L. O., Campbell Dyke, N., and Herscovics, A. (1998) Molecular cloning, chromosomal mapping and tissue-specific expression of a novel human alpha1,2-mannosidase gene involved in N-glycan maturation. Glycobiology 8, 585–595.

    Article  PubMed  CAS  Google Scholar 

  193. Zhu, A., Wang, Z. K., and Beavis, R. (1998) Structural studies of alpha-N-acetylgalactosaminidase: effect of glycosylation on the level of expression, secretion efficiency, and enzyme activity. Arch. Biochem. Biophys. 352, 1–8.

    Article  PubMed  CAS  Google Scholar 

  194. Kang, H. A., Sohn, J. H., Choi, E. S., Chung, B. H., Yu, M. H., and Rhee, S. K. (1998) Glycosylation of human alpha 1-antitrypsin in Saccharomyces cerevisiae and methylotrophic yeasts. Yeast 14, 371–381.

    Article  PubMed  CAS  Google Scholar 

  195. Lee, K. N., Tae, W.-C., Jackson, K. W., Kwon, S. H., and McKee, P. A. (1999) Characterization of wild-type and mutant alpha 2-antiplasmins: fibrinolysis enhancement by reactive site mutant. Blood 94, 164–171.

    PubMed  CAS  Google Scholar 

  196. Talmont, F., Sidobre, S., Demange, P., Milon, A., and Emorine, L. J. (1996) Expression and pharmacological characterization of the human □-opioid receptor in the methylotrophic yeast Pichia pastoris. FEBS Lett. 394, 268–272.

    Article  PubMed  CAS  Google Scholar 

  197. O’Connell, M. A., Gerber, A., and Keegan, L. P. (1998) Purification of native and recombinant double-stranded RNA-specific adenosine deaminases. Methods 15, 51–62.

    Article  PubMed  CAS  Google Scholar 

  198. Le Brocque, D., Henry, A., Cappai, R., Li, Q. X., Tanner, J. E., Galatis, D., Gray, C., Holmes, S., Underwood, J. R., Beyreuther, K., Masters, C. L., and Evin, G. (1998) Processing of the Alzheimer’s disease amyloid precursor protein in Pichia pastoris: immunodetection of alpha-, beta-, and gamma-secretase products. Biochemistry 37, 14958–14965.

    Article  PubMed  Google Scholar 

  199. Mok, S. S., Sberna, G., Heffernan, D., Cappai, R., Galatis, D., Clarris, H. J., Sawyer, W. H., Beyreuther, K., Masters, C. L., and Small, D. H. (1997) Expression and analysis of heparin-binding regions of the amyloid precursor protein of Alzheimer’s disease. FEBS Lett. 415, 303–307.

    Article  PubMed  CAS  Google Scholar 

  200. Cappai, R., Mok, S. S., Galatis, D., Tucker, D. F., Henry, A., Beyreuther, K., Small, D. H. and Masters, C. L. (1999) Recombinant human amyloid precursor-like protein 2 (APLP2) expressed in the yeast Pichia pastoris can stimulate neurite outgrowth. FEBS Lett. 442, 95–98.

    Article  PubMed  CAS  Google Scholar 

  201. Henry, A., Masters, C. L., Beyreuther, K., and Cappai, R. (1997) Expression of human amyloid precursor protein ectodomains in Pichia pastoris: analysis of culture conditions, purification, and characterization. Protein Expr. Purif. 10, 283–291.

    Article  PubMed  CAS  Google Scholar 

  202. Culvenor, J. G., Henry, A., Hartmann, T., Evin, G., Galatis, D., Friedhuber, A., Jayasena, U. L., Underwood, J. R., Beyreuther, K., Masters, C. L., and Cappai, R. (1998) Subcellular localization of the Alzheimer’s disease amyloid precursor protein and derived polypeptides expressed in a recombinant yeast system. Amyloid 5, 79–89.

    PubMed  CAS  Google Scholar 

  203. Markaryan, A., Morosova, I., Lee, B.-S., and Kaplan, A. (1999) Atypical processing of amyloid precursor fusion protein by proteolytic activity in Pichia pastoris. Biochem. Biophys. Res. Commun. 262, 263–268.

    Article  PubMed  CAS  Google Scholar 

  204. Ohsawa, I., Hirose, Y., Ishiguro, M., Imai, Y., Ishiura, S., and Kohsaka, S. (1995) Expression, purification, and neurotrophic activity of amyloid precursor protein-secreted forms produced by yeast. Biochem. Biophys. Res. Commun. 213, 52–58.

    Article  PubMed  CAS  Google Scholar 

  205. Sahasrabudhe, A. V., Solapure, S. M., Khurana, R., Suryanarayan, V., Ravishankar, S., deSousa, S. M., and Das, G. (1998) Production of recombinant human bile salt stimulated lipase and its variant in Pichia pastoris. Protein Expr. Purif. 14, 425–433.

    Article  PubMed  CAS  Google Scholar 

  206. FitzGerald, K., Hollinger, P., and Winter, G. (1997) Improved tumour targeting by disulphide stabilized diabodies expressed in Pichia pastoris. Protein Eng. 10, 1221–1225.

    Article  PubMed  CAS  Google Scholar 

  207. Lam, L. P. and Berger, S. A. (1997) Intracellular expression and purification of the c-kit receptor kinase domain in Pichia pastoris. Biotechniques 23, 83–86.

    Google Scholar 

  208. Lam, L. P., Chow, R. Y., and Berger, S. A. (1999) A transforming mutation enhances the activity of the c-Kit soluble tyrosine kinase domain. Biochem. J. 338, 131–138.

    Article  PubMed  CAS  Google Scholar 

  209. You, Y. H., Hefta, L. J., Yazaki, P. J., Wu, A. M., and Shively, J. E. (1998) Expression, purification, and characterization of a two domain carcinoembryonic antigen minigene (N-A3) in Pichia pastoris. The essential role of the N-domain. Anticancer Res. 18, 3193–3201.

    PubMed  CAS  Google Scholar 

  210. Sun, J., Bottomley, S. P., Kumar, S., and Bird, P. I. (1997) Recombinant caspase-3 expressed in Pichia pastoris is fully activated and kinetically indistinguishable from the native enzyme. Biochem. Biophys. Res. Commun. 238, 920–924.

    Article  PubMed  CAS  Google Scholar 

  211. Linnevers, C. J., McGrath, M. E., Armstrong, R., Mistry, F. R., Barnes, M. G., Klaus, J. L., Palmer, J. T., Katz, B. A., and Bromme, D. (1997) Expression of human cathepsin K in Pichia pastoris and preliminary crystallographic studies of an inhibitor complex. Protein Sci. 6, 919–921.

    Article  PubMed  CAS  Google Scholar 

  212. Hou, W. S., Brommer, D., Zhao, Y., Mehler, E., Dushey, C., Weinstein, H., Miranda, C. S., Fraga, C., Greig, F., Carey, J., Rimoin, D. L., Desnick, R. J., and Gelb, B. D. (1999) Characterization of novel cathepsin K mutations in the pro and mature polypeptide regions causing pycnodysostosis. J. Clin. Invest. 103, 731–738.

    PubMed  CAS  Google Scholar 

  213. Menard, R., Carmona, E., Takebe, S., Dufour, E., Plouffe, C., Mason, P., and Mort, J. S. (1998) Autocatalytic processing of recombinant human procathepsin L. Contribution of both intermolecular and unimolecular events in the processing of procathepsin L in vitro. J. Biol. Chem. 273, 4478–4484.

    Article  PubMed  CAS  Google Scholar 

  214. Carmona, E., Dufour, E., Plouffe, C., Takebe, S., Mason, P., Mort, J. S., and Menard, R. (1996) Potency and selectivity of the cathepsin L propeptide as an inhibitor of cysteine proteases. Biochemistry 35, 8149–8157.

    Article  PubMed  CAS  Google Scholar 

  215. Bromme, D., Li, Z., Barnes, M., and Mehler, E. (1999) Human cathepsin V functional expression, tissue distribution, electrostatic surface potential, enzymatic characterization, and chromosomal localization. Biochemistry 38, 2377–2385.

    Article  PubMed  CAS  Google Scholar 

  216. Nägler, D. K., Zhang, R., Tam, W., Sulea, T., Purisima, E. O., and Menard, R. (1999) Human cathepsin X: a cysteine protease with unique carboxypeptidase activity. Biochemistry 38, 12648–12654.

    Article  PubMed  CAS  Google Scholar 

  217. Munshi, C. B., Fryxell, K. B., Lee, H. C., and Branton, W. D. (1997) Large-scale production of human CD38 in yeast by fermentation. Methods Enzymol. 280, 318–330.

    Article  PubMed  CAS  Google Scholar 

  218. McGrew, J. T., Leiske, D., Dell, B., Klinke, R., Krasts, D., Wee, S. F., Abbott, N., Armitage, R., and Harrington, K. (1997) Expression of trimeric CD40 ligand in Pichia pastoris: use of a rapid method to detect high-level expressing transformants. Gene 187, 193–200.

    Article  PubMed  CAS  Google Scholar 

  219. Gerstmayer, B., Altenschmidt, U., Hoffmann, M., and Wels, W. (1997) Costimulation of T cell proliferation by a chimeric B7-2 antibody fusion protein specifically targeted to cells expressing the erbB2 proto-oncogene. J. Immunol. 158, 4584–4590.

    PubMed  CAS  Google Scholar 

  220. Sen Gupta, C. and Dighe, R. R. (1999) Hyperexpression of biologically active human chorionic gonadotropin using the methylotropic yeast Pichia pastoris. J. Mol. Endocrinol. 22, 273–283.

    Article  PubMed  CAS  Google Scholar 

  221. Daniels, G. L., Green, C. A., Powell, R. M., and Ward, T. (1998) Hemagglutination inhibition of Cromer blood group antibodies with soluble recombinant decay-accelerating factor. Transfusion 38, 332–336.

    Article  PubMed  CAS  Google Scholar 

  222. Battista, M. C., Bergamini, G., Campanini, F., Landini, M. P., and Ripalti, A. (1996) Intracellular production of a major cytomegalovirus antigenic protein in the methylotrophic yeast Pichia pastoris. Gene 176, 197–201.

    Article  PubMed  CAS  Google Scholar 

  223. Powell, R. M., Ward, T., Evans, D. J., and Almond, J. W. (1997) Interaction between echovirus 7 and its receptor, decay-accelerating factor (CD55): evidence for a secondary cellular factor in A-particle formation. J. Virol. 71, 9306–9312.

    PubMed  CAS  Google Scholar 

  224. Gerber, A., O’Connell, M. A., and Keller, W. (1997) Two forms of human double-stranded RNA-specific editase 1 (hRED1) generated by the insertion of an Alu cassette. RNA 3, 453–463.

    PubMed  CAS  Google Scholar 

  225. Boehm, T., O’Reilly, M. S., Keough, K., Shiloach, J., Shapiro, R., and Folkman, J. (1998) Zinc-binding of endostatin is essential for its angiogenic activity. Biochem. Biophys. Res. Commun. 252, 190–194.

    Article  PubMed  CAS  Google Scholar 

  226. Zhan, H., Liu, B., Reid, S. W., Aoki, K. H., Li, C., Syed, R. S., Karkaria, C., Koe, G., Sitney, K., Hayenga, K., Mistry, F., Savel, L., Dreyer, M., Katz, B. A., Schreurs, J., Matthews, D. J., Cheetham, J. C., Egrie, J., Giebel, L., and Stroud, R. M. (1999) Engineering a soluble extracellular erythropoietin receptor (EPObp) in Pichia pastoris to eliminate microheterogeneity, and its complex with erythropoietin. Protein Eng. 12, 505–513.

    Article  PubMed  CAS  Google Scholar 

  227. Tanaka, M., Suda, T., Yatomi, T., Nakamura, N., and Nagata, S. (1997) Lethal effect of recombinant human Fas ligand in mice pretreated with Propionibacterium acnes. J. Immunol. 158, 2303–2309.

    PubMed  CAS  Google Scholar 

  228. Cote, H. C., Pratt, K. P., Davie, E. W., and Chung, D. W. (1997) The polymerization pocket “a” within the carboxy-terminal region of the gamma chain of human fibrinogen is adjacent to but independent from the calcium-binding site. J. Biol. Chem. 272, 23792–23798.

    Article  PubMed  CAS  Google Scholar 

  229. Rosenfeld, S. A., Ross, O. H., Hillman, M. C., Corman, J. I., and Dowling, R. L. (1996) Production and purification of human fibroblast collagenase (MMP-1) expressed in the methylotrophic yeast Pichia pastoris. Protein Expr. Purif. 7, 423–430.

    Article  PubMed  CAS  Google Scholar 

  230. Spraggon, G., Applegate, D., Everse, S. J., Zhang, J. Z., Veerapandian, L., Redman, C., Doolittle, R. F., and Grieninger, G. (1998) Crystal structure of a recombinant alpha EC domain from human fibrinogen-420. Proc. Natl. Acad. Sci. U.S.A. 95, 9099–9104.

    Article  PubMed  CAS  Google Scholar 

  231. Yamada, M., Azuma, T., Matsuba, T., Iida, H., Suzuki, H., Yamamoto, K., Kohli, Y., and Hori, H. (1994) Secretion of human intracellular aspartic proteinase cathepsin E expressed in the methylotrophic yeast, Pichia pastoris and characterization of produced recombinant cathepsin E. Biochim. Biophys. Acta 1206, 279–285.

    PubMed  CAS  Google Scholar 

  232. Roy, N., Padmanabhan, S., Smith, M., Shi, L., Navre, M., and Das, G. (1999) Expression of human gelatinase B in Pichia pastoris. Protein Expr. Purif. 16, 324–330.

    Article  PubMed  CAS  Google Scholar 

  233. Sun, J., Bird, C. H., Buzza, M. S., McKee, K. E., Whisstock, J. C., and Bird, P. I. (1999) Expression and purification of recombinant human granzyme B from Pichia pastoris. Biochem. Biophys. Res. Commun. 261, 251–255.

    Article  PubMed  CAS  Google Scholar 

  234. Zhu, H., Shi, J., Cregg, J. M., and Woldegiorgis, G. (1997) Reconstitution of highly expressed human heart muscle carnitine palitoyltransferase I. Biochem. Biophys. Res. Commun. 239, 498–502.

    Article  PubMed  CAS  Google Scholar 

  235. Kjeldsen, T., Pettersson, A. F., and Hach, M. (1999) Secretory expression and characterization of insulin in Pichia pastoris. Biotechnol. Appl. Biochem. 29, 79–86.

    PubMed  CAS  Google Scholar 

  236. Brierley, R. A. (1998) Secretion of recombinant human insulin-like growth factor I (IGF-1). Methods Mol. Biol. 103, 149–177.

    PubMed  CAS  Google Scholar 

  237. Green, M. M., Isarkin, J., III, Subramaniam, P. S., Szente, B. E., and Johnson, H. M. (1998) Human IFN gamma receptor cytoplasmic domain: expression and interaction with HuIFN gamma. Biochem. Biophys. Res. Commun. 243, 170–176.

    Article  PubMed  CAS  Google Scholar 

  238. Johnson, T. M., Holaday, S. K., Sun, Y., Subramaniam, P. S., Johnson, H. M., and Krishna, N. R. (1999) Expression, purification, and characterization of interferon-tau produced in Pichia pastoris grown in a minimal medium. J. Interferon Cytokine Res. 19, 631–636.

    Article  PubMed  CAS  Google Scholar 

  239. Murphy, K. P., Jr., Gagne, P., Pazmany, C., and Moody, M. D. (1998) Expression of human interleukin-17 in Pichia pastoris: purification and characterization. Protein Expr. Purif. 12, 208–214.

    Article  PubMed  CAS  Google Scholar 

  240. Sun, J., Coughlin, P., Salem, H. H., and Bird, P. (1995) Production and characterization of recombinant human proteinase inhibitor 6 expressed in Pichia pastoris. Biochim. Biophys. Acta 1252, 28–34.

    PubMed  Google Scholar 

  241. Wagner, S. L., Siegel, R. S., Vedvick, T. S., Raschke, W. C., and Van Nostrand, W. E. (1992) High level expression, purification, and characterization of the Kunitz-type protease inhibitor domain of protease nexin-2/amyloid beta-protein precursor. Biochem. Biophys. Res. Commun. 186, 1138–1145.

    Article  PubMed  CAS  Google Scholar 

  242. Zhang, J. G., Owczarek, C. M., Ward, L. D., Howlett, G. J., Fabri, L. J., Roberts, B. A., and Nicola, N. A. (1997) Evidence for the formation of a heterotrimeric complex of leukaemia inhibitory factor with its receptor subunits in solution. Biochem. J. 325, 693–700.

    PubMed  CAS  Google Scholar 

  243. Fryxell, K. B., O’Donoghue, K., Graeff, R. M., Lee, H. C., and Branton, W. D. (1995) Functional expression of soluble forms of human CD38 in Escherichia coli and Pichia pastoris. Protein Expr. Purif. 6, 329–336.

    Article  PubMed  CAS  Google Scholar 

  244. Liao, Y. F., Lal, A., and Moremen, K. W. (1996) Cloning, expression, purification, and characterization of the human broad specificity lysosomal acid alpha-mannosidase. J. Biol. Chem. 271, 28348–28358.

    Article  PubMed  CAS  Google Scholar 

  245. Chan, H., Elrod, K. C., Numerof, R. P., Sideris, S., and Clark, J. M. (1999) Expression and characterization of recombinant mast cell tryptase. Protein Expr. Purif. 15, 251–257.

    Article  PubMed  CAS  Google Scholar 

  246. Niles, A. L., Maffitt, M., Haak-Frendscho, M., Wheeless, C. J., and Johnson, D. A. (1998) Recombinant human mast cell tryptase beta: stable expression in Pichia pastoris and purification of fully active enzyme. Biotechnol. Appl. Biochem. 28, 125–131.

    PubMed  CAS  Google Scholar 

  247. Kalandadze, A., Galleno, M., Foncerrada, L., Strominger, J. L., and Wucherpfennig, K. W. (1996) Expression of recombinant HLA-DR2 molecules. Replacement of the hydrophobic transmembrane region by a leucine zipper dimerization motif allows the assembly and secretion of soluble DR alpha-beta heterodimers. J. Biol. Chem. 271, 20156–20162.

    Article  PubMed  CAS  Google Scholar 

  248. Luo, D., Mah, N., Krantz, M., Wilde, K., Wishart, D., Zhang, Y., Jacobs, F., and Martin, L. (1995) Vl-linker-Vh orientation-dependent expression of single chain Fv-containing an engineered disulfide-stabilized bond in the framework regions. J. Biochem. (Tokyo) 118, 825–831.

    CAS  Google Scholar 

  249. Beall, C. J., Breckenridge, S. M., Chakravarty, L., and Kolattukudy, P. E. (1998) Expression of human monocyte chemoattractant protein-1 in the yeast Pichia pastoris. Protein Expr. Purif. 12, 145–150.

    Article  PubMed  CAS  Google Scholar 

  250. Masure, S., Paemen, L., Proost, P., Van Damme, J., and Opdenakker, G. (1995) Expression of a human mutant monocyte chemotactic protein 3 in Pichia pastoris and characterization as an MCP-3 receptor antagonist. J. Interferon Cytokine Res. 15, 955–963.

    Article  PubMed  CAS  Google Scholar 

  251. Kroll, K. A., Otte, S., Hirschfeld, G., Barnikol-Watanabe, S., Gotz, H., Sternbach, H., Kratzin, H. D., Barnikol, H. U., and Hilschmann, N. (1999) Heterologous overexpression of human NEFA and studies on the two EF-hand calcium-binding sites. Biochem. Biophys. Res. Commun. 260, 1–8.

    Article  PubMed  CAS  Google Scholar 

  252. Kiselyov, V. V., Berezin, V., Maar, T. E., Soroka, V., Edvardsen, K., Schousboe, A., and Bock, E. (1997) The first immunoglobulin-like neural cell adhesion molecule (NCAM) domain is involved in double-reciprocal interaction with the second immunoglobulin-like NCAM domain and in heparin binding. J. Biol. Chem. 272, 10125–10134.

    Article  PubMed  CAS  Google Scholar 

  253. Yang, Y. S., Yang, M. C., Tucker, P. W., and Capra, J. D. (1997) NonO enhances the association of many DNA-binding proteins to their targets. Nucleic Acids Res. 25, 2284–2292.

    Article  PubMed  CAS  Google Scholar 

  254. Moritz, R. L., Ward, L. D., Tu, G.-F., Fabri, L. J., Ji, H., Yasukawa, K., and Simpson, R. J. (1999) The N-terminus of gp130 is critical for the formation of the high-affinity interleukin-6 receptor complex. Growth Factors 16, 265–278.

    Article  PubMed  CAS  Google Scholar 

  255. Rydberg, E. H., Sidhu, G., Vo, H. C., Hewitt, J., Cote, H. C., Wang, Y., Numao, S., MacGillivray, R. T., Overall, C. M., Brayer, G. D., and Withers, S. G. (1999) Cloning, mutagenesis, and structural analysis of human pancreatic alpha-amylase expressed in Pichia pastoris. Protein Sci. 8, 635–643.

    Article  PubMed  CAS  Google Scholar 

  256. Yang, Y. and Lowe, M.E. (1998) Human pancreatic triglyceride lipase expressed in yeast cells: purification and characterization. Protein Expr. Purif. 13, 36–40.

    Article  PubMed  CAS  Google Scholar 

  257. Dufour, E., Tam, W., Nagler, D. K., Storer, A. C., and Menard, R. (1998) Synthesis of amidrazones using an engineered papain nitrile hydratase. FEBS Lett. 433, 78–82.

    Article  PubMed  CAS  Google Scholar 

  258. Heimo, H., Palmu, K., and Suominen, I. (1998) Human placental alkaline phosphatase: expression in Pichia pastoris, purification and characterization of the enzyme. Protein Expr. Purif. 12, 85–92.

    Article  PubMed  CAS  Google Scholar 

  259. Dutta, B., Mukhopadhyay, D., Roy, N., Das, G., and Karande, A. A. (1998) Cloning, expression purification, and immunocharacterization of placental protein-14. Protein Expr. Purif. 14, 327–334.

    Article  PubMed  CAS  Google Scholar 

  260. Duman, J. G., Miele, R. G., Liang, H., Grella, D. K., Sim, K. L., Castellino, F. J., and Bretthauer, R. K. (1998) O-Mannosylation of Pichia pastoris cellular and recombinant proteins. Biotechnol. Appl. Biochem. 28, 39–45.

    PubMed  CAS  Google Scholar 

  261. Sim, B. K., O’Reilly, M. S., Liang, H., Fortier, A. H., He, W., Madsen, J. W., Lapcevich, R., and Nacy, C. A. (1997) A recombinant human angiostatin protein inhibits experimental primary and metastatic cancer. Cancer Res. 57, 1329–1334.

    PubMed  CAS  Google Scholar 

  262. Nilsen S. L., Prorok, M., Castellino, F. J. (1999) Enhancement through mutagenesis of the binding of the isolated kringle 2 domain of human plasminogen to omega-amino acid ligands and to an internal sequence of a Streptococcal surface protein. J. Biol. Chem. 274, 22380–22386.

    Article  PubMed  CAS  Google Scholar 

  263. Reverter, D., Ventura, S., Villegas, V., Vendrell, J., and Aviles, F. X. (1998) Overexpression of human procarboxypeptidase A2 in Pichia pastoris and detailed characterization of its activation pathway. J. Biol. Chem. 273, 3535–3541.

    Article  PubMed  CAS  Google Scholar 

  264. Illy, C., Quraishi, O., Wang, J., Purisima, E., Vernet, T., and Mort, J. S. (1997) Role of occluding loop in cathepsin B activity. J. Biol. Chem. 272, 1197–1202.

    Article  PubMed  CAS  Google Scholar 

  265. Cordle, R. A. and Lowe, M. E. (1998) Purification and characterization of human procolipase expressed in yeast cells. Protein Expr. Purif. 13, 30–35.

    Article  PubMed  CAS  Google Scholar 

  266. Lima, C. D., Klein, M. G., Weinstein, I. B., and Hendrickson, W. A. (1996) Three-dimensional structure of human protein kinase C interacting protein 1, a member of the HIT family of proteins. Proc. Natl. Acad. Sci. U.S.A. 93, 5357–5362.

    Article  PubMed  CAS  Google Scholar 

  267. Harmsen, M. C., Heeringa, P., van der Geld, Y. M., Huitema, M. G., Klimp, A., Tiran, A., and Kallenberg, C. G. (1997) Recombinant proteinase 3 (Wegener’s antigen) expressed in Pichia pastoris is functionally active and is recognized by patient sera. Clin. Exp. Immunol. 110, 257–264.

    Article  PubMed  CAS  Google Scholar 

  268. Dahlen, J. R., Foster, D. C., and Kisiel, W. (1997) Expression, purfication, and inhibitory properties of human proteinase inhibitor. Biochemistry 36, 14874–14882.

    Article  CAS  Google Scholar 

  269. Luo, D., Geng, M., Schultes, B., Ma, J., Xu, D. Z., Hamza, N., Qi, W., Noujaim, A. A., and Madiyalakan, R. (1998) Expression of a fusion protein of scFv-biotin mimetic peptide for immunoassay. J. Biotechnol. 65, 225–228.

    Article  PubMed  CAS  Google Scholar 

  270. Pennell, C. A. and Eldin, P. (1998) In vitro production of recombinant antibody fragments in Pichia pastoris. Res. Immunol. 149, 599–603.

    Article  PubMed  CAS  Google Scholar 

  271. Ohtani, W., Nawa, Y., Takeshima, K., Kamuro, H., Kobayashi, K., and Ohmura, T. (1998) Physicochemical and immunochemical properties of recombinant human serum albumin from Pichia pastoris. Anal. Biochem. 256, 56–62.

    Article  PubMed  CAS  Google Scholar 

  272. Ikegaya, K., Hirase, M., Ohmura, T., and Nokihara, K. (1997) Complete determination of disulfide forms of purified recombinant human serum albumin, secreted by the yeast Pichia pastoris. Anal. Chem. 69, 1986–1991.

    Article  PubMed  CAS  Google Scholar 

  273. Ohtani, W., Ohda, T., Sumi, A., Kobayashi, K., and Ohmura, T. (1998) Analysis of Pichia pastoris components in recombinant human serum albumin by immunological assays and by HPLC with pulsed amperometric detection. Anal. Chem. 70, 425–429.

    Article  PubMed  CAS  Google Scholar 

  274. Petersen, C. E., Ha, C.-E., Harohalli, K., Park, D. S., Feix, J. B., Isozaki, O., and Bhagavan, N. V. (1999) Structural investigations of a new familial dysalbuminemic hyperthyroxinemia genotype. Clin. Chem. 45, 1248–1254.

    PubMed  CAS  Google Scholar 

  275. Steinlein, L. M., Graf, T. N., and Ikeda, R. A. (1995) Production and purification of N-terminal half-transferrin in Pichia pastoris. Protein Expr. Purif. 6, 619–624.

    Article  PubMed  CAS  Google Scholar 

  276. Bewley, M. C., Tam, B. M., Grewal, J., He, S., Shewry, S., Murphy, M. E., Mason, A. B., Woodworth, R. C., Baker, E. N., and MacGillivray, R. T. (1999) X-ray crystallography and mass spectroscopy reveal that the N-lobe of human transferrin expressed in Pichia pastoris is folded correctly but is glycosylated on serine-32. Biochemistry 38, 2535–2541.

    Article  PubMed  CAS  Google Scholar 

  277. Mason, A. B., Woodworth, R. C., Oliver, R. W. A., Green, B. N., Lin, L. N., Brandts, J. F., Tam, B. M., Maxwell, A., and MacGillivray, R. T. (1996) Production and isolation of the recombinant N-lobe of human serum transferrin from the methylotrophic yeast Pichia pastoris. Protein Expr. Purif. 8, 119–125.

    Article  PubMed  CAS  Google Scholar 

  278. Sui, L. M., Lennon, J., Ma, C., McCann, I., Woo, I., and Petra, P. H. (1999) Heterologous expression of wild type and deglycosylated human sex steroid-binding protein (SBP or SHBG) in the yeast, Pichia pastoris. Characterization of the recombinant proteins. J. Steroid Biochem. Mol. Biol. 68, 119–127.

    Article  PubMed  CAS  Google Scholar 

  279. Tsujikawa, M., Okabayashi, K., Morita, M., and Tanabe, T. (1996) Secretion of a variant of human single-chain urokinase-type plasminogen activator without an N-glycosylation site in the methylotrophic yeast, Pichia pastoris and characterization of the secreted product. Yeast 12, 541–553.

    Article  PubMed  CAS  Google Scholar 

  280. White, C. E., Hunter, M. J., Meininger, D. P., White, L. R., and Komives, E. A. (1995) Large-scale expression, purification and characterization of small fragments of thrombomodulin: the roles of the sixth domain and of methionine 388. Protein Eng. 8, 1177–1187.

    Article  PubMed  CAS  Google Scholar 

  281. Austin, A. J., Jones, C. E., and Heeke, G. V. (1998) Production of human tissue factor using the Pichia pastoris expression system. Protein Expr. Purif. 13, 136–142.

    Article  PubMed  CAS  Google Scholar 

  282. Chan, H., Springmann, E. B., and Clark, J. M. (1998) Expression and characterization of human tissue kallikrein variants. Protein Expr. Purif. 12, 361–370.

    Article  PubMed  CAS  Google Scholar 

  283. Katz, B. A., Liu, B., Barnes, M., and Springman, E. B. (1998) Crystal structure of recombinant human tissue kallikroin at 2.0 Å resolution. Protein Sci. 7, 875–885.

    Article  PubMed  CAS  Google Scholar 

  284. Cregg, J. M. and Madden, K. R. (1989) Use of site-specific recombination to regenerate selectable markers. Mol. Gen. Genet. 219, 320–323.

    Article  PubMed  CAS  Google Scholar 

  285. Miele, R. G., Castellino, F. J., and Bretthauer, R. K. (1997) Characterization of the acidic oligosaccharides assembled on the Pichia pastoris-expressed recombinant kringle 2 domain of human tissue-type plasminogen activator. Biotechnol. Appl. Biochem. 26, 79–83.

    PubMed  CAS  Google Scholar 

  286. Miele, R. G., Nilsen, S. L., Brito, T., Bretthauer, R. K., and Castellino, F. J. (1997) Glycosylation properties of the Pichia pastoris-expressed recombinant kringle 2 domain of tissue-type plasminogen activator. Biotechnol. Appl. Biochem. 25, 151–157.

    PubMed  CAS  Google Scholar 

  287. Chang, T., Zajicek, J., and Castellino, F. J. (1997) Role of tryptophan-63 of the kringle 2 domain of tissue-type plasminogen activator in its thermal stability, folding, and ligand binding properties. Biochemistry 36, 7652–7663.

    Article  PubMed  CAS  Google Scholar 

  288. Glansbeek, H. L., van Beuningen, H. M., Vitters, E. L., van der Kraan, P. M., and van den Berg, W. B. (1998) Expression of recombinant human soluble type II transforming growth factor-beta receptor in Pichia pastoris and Escherichia coli: two powerful systems to express a potent inhibitor of transforming growth factor-beta. Protein Expr. Purif. 12, 201–207.

    Article  PubMed  CAS  Google Scholar 

  289. Sreekrishna, K., Potenz, R. H. B., Cruze, J. A., McCombie, W. R., Parker, K. A., Nelles, L., Mazzaferro, P. K., Holden, K. A., Harrison, R. G., Wood, P. J., Phelps, D. A., Hubbard, C. E., and Fuke, M. (1988) High level expression of heterologous proteins in methylotrophic yeast Pichia pastoris. J. Basic Microbiol. 28, 265–278.

    Article  PubMed  CAS  Google Scholar 

  290. Rodenburg, K. W., Kjoller, L., Petersen, H. H., and Andreasen, P. A. (1998) Binding of urokinase-type plasminogen activator-plasminogen activator inhibitor-1 complex to the endocytosis receptors alpha 2-macroglobulin receptor/low-density lipoprotein receptor-related protein and very-low-density lipoprotein receptor involves basic residues in the inhibitor. Biochem. J. 329, 55–63.

    PubMed  CAS  Google Scholar 

  291. Vuorela, A., Myllyharju, J., Nissi, R., Pihlajaniemi, T., and Kivirikko, K. I. (1997) Assembly of human prolyl 4-hydroxylase and type III collagen in the yeast Pichia pastoris: formation of a stable enzyme tetramer requires coexpression with collagen and assembly of a stable collagen requires coexpression with prolyl 4-hydroxylase. EMBO J. 16, 6702–6712.

    Article  PubMed  CAS  Google Scholar 

  292. Okabayashi, K., Tsujikawa, M., Morita, M., Einaga, K., Tanaka, K., Tanabe, T., Yamanouchi, K., Hirama, M., Tait, J. F., and Fujikawa, K. (1996) Secretory production of recombinant urokinase-type plasminogen activator-annexin V chimeras in Pichia pastoris. Gene 177, 69–76.

    Article  PubMed  CAS  Google Scholar 

  293. Mohanraj, D., Olson, T., and Ramakrishnan, S. (1995) Expression of biologically active human vascular endothelial growth factor in yeast. Growth Factors 12, 17–27.

    PubMed  CAS  Google Scholar 

  294. Saelens, X., Vanlandschoot, P., Martinet, W., Maras, M., Neirynck, S., Contreras, R., Fiers, W., and Jou, W. M. (1999) Protection of mice against a lethal influenza virus challenge after immunization with yeast-derived secreted influenza virus hemagglutinin. Eur. J. Biochem. 260, 166–175.

    Article  PubMed  CAS  Google Scholar 

  295. Martinet, W., Saelens, X., Deroo, T., Neirynck, S., Contreras, R., Min Jou, W., and Fiers, W. (1997) Protection of mice against a lethal influenza challenge by immunization with yeast-derived recombinant influenza neuraminidase. Eur. J. Biochem. 247, 332–338.

    Article  PubMed  CAS  Google Scholar 

  296. Zhu, X., Wu, S., and Letchworth, G. J., III (1997) Yeast-secreted bovine herpesvirus type 1 glycoprotein D has authentic conformational structure and immunogenecity. Vaccine 15, 679–688.

    Article  PubMed  CAS  Google Scholar 

  297. Zhu, X., Wu, S., and Letchworth, G. J. (1999) A chimeric protein comprised of bovine herpesvirus type 1 glycoprotein D and bovine interleukin-6 is secreted by yeast and possesses biological activities of both molecules. Vaccine 17, 269–282.

    Article  PubMed  CAS  Google Scholar 

  298. Sugrue, R. J., Fu, J., Howe, J., and Chan, Y. C. (1997) Expression of the dengue virus structural proteins in Pichia pastoris leads to the generation of virus-like particles. J. Gen. Virol. 78, 1861–1866.

    PubMed  CAS  Google Scholar 

  299. Chiou, H. L., Lee, T. S., Kuo, J., Mau, Y. C., and Ho, M. S. (1997) Altered antigenicity of “a” determinant variants of hepatitis B virus. J. Gen. Virol. 78, 2639–2645.

    PubMed  CAS  Google Scholar 

  300. Eckart, M. R. and Bussineau, C. M. (1996) Quality and authenticity of heterologous proteins synthesized in yeast. Curr. Opin. Biotechnol. 7, 525–530.

    Article  PubMed  CAS  Google Scholar 

  301. Lal, S. K., Tulasiram, P., and Jameel, S. (1997) Expression and characterization of the hepatitis E virus ORF3 protein in the methylotrophic yeast, Pichia pastoris. Gene 190, 63–67.

    Article  PubMed  CAS  Google Scholar 

  302. Scorer, C. A., Buckholz, R. G., Clare, J. J., and Romanos, M. A. (1993) The intracellular production and secretion of HIV-1 envelope protein in the methylotrophic yeast Pichia pastoris. Gene 136, 111–119.

    Article  PubMed  CAS  Google Scholar 

  303. Peng, Y. C. and Acheson, N. H. (1997) Production of active polyomavirus large T antigen in yeast Pichia pastoris. Virus Res. 49, 41–47.

    Article  PubMed  CAS  Google Scholar 

  304. Bisaillon, M., Bergeron, J., and Lemay, G. (1997) Characterization of the nucleoside triphosphate phosphohydrolase and helicase activities of the reovirus lambda 1 protein. J. Biol. Chem. 272, 18298–18303.

    Article  PubMed  CAS  Google Scholar 

  305. Bisaillon, M., Senechal, S., Bernier, L., and Lemay, G. (1999) A glycosyl hydrolase activity of mammalian reovirus sigma 1 protein can contribute to viral infection through a mucus layer. J. Mol. Biol. 286, 759–773.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Keck Graduate Institute of Applied Life Sciences, 535 North Watson Dr., 91711, Claremont, CA

    James M. Cregg

  2. Oregon Graduate Institute of Science and Technology, 20,000 N.W. Walker Rd. Beaverton, 97006, OR

    Joan Lin Cereghino & Jianying Shi

  3. Idun Pharmaceuticals, 11085 N., 92037, Torrey Pines Road La Jolla, CA

    David R. Higgins

Authors
  1. James M. Cregg
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Joan Lin Cereghino
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jianying Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. David R. Higgins
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to David R. Higgins.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cregg, J.M., Cereghino, J.L., Shi, J. et al. Recombinant protein expression in Pichia pastoris . Mol Biotechnol 16, 23–52 (2000). https://doi.org/10.1385/MB:16:1:23

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1385/MB:16:1:23

Index Entries

Search

Navigation