Abstract
Yeasts are attractive hosts for the production of heterologous proteins. Unlike prokaryotic systems, their eukaryotic subcellular organization enables them to carry out many of the post–translational folding, processing and modification events required to produce “authentic” and bioactive mammalian proteins. In addition, they retain the advantages of a unicellular microorganism, with respect to rapid growth and ease of genetic manipulation. The vast majority of yeast expression work has focused on the well–characterized baker's yeast Saccharomyces cerevisiae. However, with the development of DNA transformation technologies, a growing number of non–Saccharomyces yeasts are becoming available as hosts for recombinant polypeptide production. These include Hansenula polymorpha, Kluyveromyces lactis, Pichia pastoris, Schizosaccharomyces pombe, Schwanniomyces occidentalis and Yarrowia lipolytica. The performance of these alternative yeast expression systems is reviewed here relative to S. cerevisiae, and the advantages and limitations of these systems are discussed.
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References
- 1.
Itakura, K., Hirose, T., Crea, R., Riggs, A.D., Heynecker, H., Bolivar, F. and Boyer, H.W. 1977. Expression in Escherichia coli of a chemically synthesized gene for the hormone somatostatin. Science 198: 1056–1063.
- 2.
Schekman, R. and Novick, P. 1982. The secretory process and yeast cell-surface assembly, p. 361–393 In: The Molecular Biology of the Yeast Saccharomyces. Strathern, J. N., Jones, E. W., and Broach, J. R. (Eds.). Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
- 3.
Hitzeman, R.A., Hagie, F.E., Levine, H.L., Goeddel, D.V., Ammerer, G. and Hall, B.D. 1981. Expression of a human gene for interferon in yeast. Nature 293: 717–722.
- 4.
Valenzuela, P., Medina, A., Rutter, W.J., Ammerer, G. and Hall, B.D. 1982. Synthesis and assembly of hepatitis B virus surface antigen particles in yeast. Nature 298: 347–350.
- 5.
Genetic Engineering News, August, 1990.
- 6.
Hallewell, R.A., Mills, R., Tekamp-Olson, P., Blacher, R., Rosenberg, S., Otting, F., Masiarz, F.R. and Scandella, C.J. 1987. Amino terminal acetylation of authentic human Cu, Zn superoxide dismutase pro duced in yeast. Bio/Technology 5: 363–366.
- 7.
De Baetselier, A., Vasavada, A., Dohet, P., Ha-Thi, V., De Beukelaer, M., Erpicum, T., De Clerck, L., Hanoutier, J. and Rosenberg, S. 1991. Fermentation of a yeast producing A. niger glucose oxidase: scale-up, purification and characterization of the recombinant enzyme. Bio/Technology 9: 559–561.
- 8.
Da Silva, N.A. and Bailey, J.E. 1991. Influence of plasmid origin and promoter strength in fermentations of recombinant yeast. Biotechnology and Bioengineering 37: 318–324.
- 9.
Da Silva, N.A. and Bailey, J.E. 1991. Influence of dilution rate and induction of cloned gene expression in continuous fermentations of recombinant yeast. Biotechnology and Bioengineering 37: 309–317.
- 10.
Shuster, J.R. 1989. Regulated transcriptional systems for the production of proteins in yeast: regulation by carbon source, p. 83–10108 In: Yeast Genetic Engineering. Barr, P. J., Brake, A. J., and Valenzuela, P. (Eds.). Butterworth Publishers, Stoneham, MA.
- 11.
Hinnen, A., Meyhack, B. and Heim, J. 1989. Heterologous gene expression in yeast, p. 193–214 In: Yeast Genetic Engineering. Barr, P. J., Brake, A. J., and Valenzuela, P. (Eds.). Butterworth Publishers, Stoneham, MA.
- 12.
Van Arsdell, J.N., Kwok, S., Schweichart, B.L., Ladner, M.B., Gelfand, D.H. and Innis, M.A., 1987. Cloning, characterization, and expression in Saccharomyces cerevisiae of endoglucanase I from Trichoderma reesei. Bio/Technology 5: 60–64.
- 13.
Moir, D.T. and Dumais, D.R. 1987. Glycosylation and secretion of human alpha-1-antitrypsin by yeast. Gene 56: 209–217.
- 14.
Lemontt, J.F., Wei, C.-M. and Dackowski, W.R. 1985. Expression of active human uterine tissue plasminoeen activator in yeast. DNA 4: 419–428.
- 15.
Smith, R.A., Duncan, M.J. and Moir, D.T. 1985. Heterologous protein secretion from yeast. Science 229: 1219–1224.
- 16.
Zsebo, K.M., Lu, H.-S., Fieschko, J.C., Goldstein, L., Davis, J., Duker, K., Suggs, S.V., Lai, P.H. and Bitter, G.A. 1986. Protein secretion from Saccharomyces cerevisiae directed by the prepro-α-factor leader region. J. Biol. Chem 261: 5858–5865.
- 17.
Jigami, Y., Muraki, M., Harada, N. and Tanaka, H. 1986. Expression of synthetic human-lysozyme gene in Saccharomyces cerevisiae: use of a synthetic chicken-lysozyme signal sequence for secretion and processing. Gene 43: 273–279.
- 18.
De Nobel, J.G. and Barnett, J.A. 1991. Passage of molecules through yeast cell walls: a brief essay-review. Yeast 7: 313–323.
- 19.
Reiser, J., Glumoff, V., Kalin, M. and Ochsner, U. 1990. Transfer and expression of heterologous genes in yeasts other than Saccharomyces cerevisiae. Adv. Bioch. Eng. Biotech. 43: 75–102.
- 20.
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. Bio/ Technology 5: 479–485.
- 21.
Janowicz, Z.A., Melber, K., Merckelbach, A., Jacobs, E., Harford, N., Comberbach, M. and Hollenberg, C.P. 1991. Simultaneous expression of the S and L surface antigens of hepatitis B, and formation of mixed particles in the methylotrophic yeast, Hansenula polymorpha. Yeast 7: In press.
- 22.
Siegel, R.S., Buckholz, R. and Thill, G.P. 1989. Production of epidermal growth factor in Pichia pastoris yeast cells. European Patent application W090/10697.
- 23.
Gellissen, G., Janowicz, Z.A., Merckelbach, A., Piontek, M., Keup, P., Weydemann, U., Hollenberg, C.P. and Strasser, A.W.M. 1991. Heterologous gene expression in Hansenula polymorpha: efficient secretion of glucoamylase. Bio/Technology 9: 291–295.
- 24.
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. Bio/Technology 5: 1305–1308.
- 25.
Sierkstra, L.N., Verbakel, J.M.A. and Verrips, C.T. 1991. Optimisation of a host/vector system for heterologous gene expression by Hansenula polymorpha. Curr. Genet. 19: 81–87.
- 26.
van den Berg, J.A., van der Laken, K.J., van Ooyen, A.J.J., Renniers, T.C.H.M., Rietveld, D. and Schaap, A., Brake, A.J., Bishop, R.J., Schultz, K., Moyer, D., Richman, M. and Shuster, J.R. 1990. Kluyveromyces as a host for heterologous gene expression: expression and secretion of prochymosin. Bio/Technology 8: 135–139.
- 27.
Fleer, R., Chen, X.J., Amellal, N., Yeh, P., Fournier, A., Guinet, F., Gault, N., Faucher, D., Folliard, F., Fukuhara, H. and Mayaux, J.-F. 1991. High-level secretion of correctly processed recombinant human interleukin-1β in Kluyveromyces lactis. Gene, In press.
- 28.
Fleer, R., Yeh, P., Amellal, N., Maury, I., Fournier, A., Bacchetta, F., Baduel, P., Jung, G., L'Hote, H., Becquart, J., Fukuhara, H. and Mayaux, J.F. 1991. Stable multicopy vectors for high-level secretion of recombinant human serum albumin in Kluyveromyces yeasts. Bio/ Technology 9: 968–975.
- 29.
Baldara, C., Murray, J.A.H., Ghiara, P., Cesareni, G. and Galeotti, C.L. 1987. A novel leader peptide which allows efficient secretion of a fragment of human interleukin 1 in Saccharomyces cerevisiae. EMBO J. 6: 229–234.
- 30.
Kappeli, O. 1986. Regulation of carbon metabolism in Saccharomyces cerevisiae and related yeasts. Adv. Microb. Phys. 28: 183–209.
- 31.
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.
- 32.
Ledeboer, A.M., Edens, L., Maat, J., Visser, B.J.W., Verrips, C.T., Janowicz, Z., Eckart, M., Roggenkamp, R. and Hollenberg, C.P. 1985. Molecular cloning and characterization of a gene coding for methanol oxidase in Hansenula polymorpha. Nucl. Acids Res. 13: 3063–3082.
- 33.
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. Nucl. Acids Res. 9: 3859–3876.
- 34.
Gleeson, M.A.G. and Sudbery, P.E. 1988. The methylotrophic yeasts. Yeast 4: 1–15.
- 35.
Piontek, M., Hollenberg, C.P. and Strasser, A.W.M. 1990. Schwanniomyces occidentals: a promising system for the expression of foreign genes. Yeast 6S: 422.
- 36.
Franke, A.E., Kaczmarek, F.S., Eisenhard, M.E., Geoghegan, K.F., Danley, D.E., De Zeeuw, J.R., O'Donnell, M.M., Gollaher, M.G. and Davidow, L.S. 1988. Expression and secretion of bovine prochymosin in Yarrowia lipolytica. Devel. Ind. Microbiol. 29: 43–57.
- 37.
Parent, S.A., Fenimore, C.M. and Bostian, K.A. 1985. Vector systems for the expression, analysis and cloning of DNA sequences in S. cerevisiae. Yeast 1: 83–138.
- 38.
Vedvick, T., Buckholz, R., Engel, M., Urcan, M., Kinney, J., Provow, S., Siegel, R. and Thill, G. 1991. High level secretion of biologically active aprotinin from the yeast Pichia pastoris. J. Indust. Microbiol. 7: 197–202.
- 39.
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., Velicelebi, G., Harpold, M.M. and Thill, G.P. 1989. Continuous production of a novel lysozyme via secretion from the yeast, Pichia pastoris. Bio/Technology 7: 160–164.
- 40.
Sleep, D., Belfield, G.P., Ballance, D.J., Steven, J., Jones, S., Evans, L.R., Moir, P.D. and Goodey, A.R. 1991. Saccharomyces cerevisiae strains that overexpress heterologous proteins. Bio/Technology 9: 183–187.
- 41.
Sakai, A., Shimiza, Y. and Hishinama, F. 1988. Isolation and charac terisation of mutants which show an oversecretion phenotype in Saccharomyces cerevisiae. Genetics 119: 499–506.
- 42.
Heslot, H., Nicaud, J.M., Fabre, E., Beckerich, J.M., Fournier, P. and Gaillardin, C. 1990. Cloning of the alkaline extracellular protease gene of Yarrowia lipolytica and its use to express foreign genes, p. 27–45 In: Microbiology Applications in Food Biotechnology. Nga, B. H., and Lee, Y. K. (Eds.). Elsevier Applied Science, New York, NY.
- 43.
Davidow, L.S., Franke, A.E. and De Zeeuw, J.R. 1987. New Yarrowia lipolytica transformants used for expression and secretion of heterologous proteins, especially prorennin and human anaphylatoxin C5a. European Patent Application EP86307839.
- 44.
Bröker, M. and Bauml, O. 1989. New expression vectors for the fission yeast Schizosaccharomyces pombe. FEES Lett. 248: 105.
- 45.
Grinna, L.S. and Tschopp, J.F. 1989. Size distribution and general structural features of N-linked oligosaccharides from the methylo-trophic yeast, Pichia pastoris. Yeast 5: 107–115.
- 46.
Tarentino, A.L., Plummer, T.H. and Maley, F. 1974. The release of intact oligosaccharides from specific glycoproteins by endo—N-acetyl-glucosaminidase H. J. Biol. Chem. 249: 818–824.
- 47.
Esmon, B., Esmon, P.C. and Schekman, R. 1984. Early steps in processing of yeast glycoproteins. J. Biol. Chem 259: 10322–10327.
- 48.
Deibel, M.R., Hiebsch, R.R. and Klein, R.D. 1988. Secreted amylolytic enzymes from Schwanniomyces occidentalis: purification by fast protein liquid chromatography (FPLC) and preliminary characterization. Prep. Biochem. 18: 77–120
- 49.
Julius, D., Brake, A., Blair, L., Kunisawa, R. and Thorner, J. 1984. Isolation of the putative structural gene for the lysine-arginine-cleaving endopeptidase required for processing of yeast prepro-α-factor. Cell 37: 1075–1089.
- 50.
Hodgkins, M.A., Sudbery, P.E., Perry-Williams, S. and Goodey, A. 1990. Secretion of human serum albumin from Hansenula polymorpha. Yeast 6S: 435.
- 51.
Brake, A., Irvine, B., Masiarz, F. and Schultz, K. 1988. Structure of genes encoding precursors of two Kluyveromyces lactis transported proteins. Yeast 4S: 436.
- 52.
Tanguy-Rougeau, C., Wesoslowski-Louvel, M. and Fukuhara, H., 1988. The Kluyveromyces lactis KEX1 gene encodes a subtilisin-type serine proteinase. Yeast 4S: 186.
- 53.
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
- 54.
Clare, J.J., Rayment, F.B., Ballantine, S.P., Sreekrishna, K. and Romanes, M.A. 1991. High-level expression of tetanus toxin frag ment C in Pichia pastoris strains containing multiple tandem integations of the gene. Bio/Technology 9: 455–460.
- 55.
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.
- 56.
Shen, S.H., Bastien, L., Nguyen, T., Fung, M. and Slilaty, S.N. 1989. Synthesis and secretion of hepatitis B middle surface antigen by the methylotrophic yeast, Hansenula polymorpha. Gene 84: 303–309.
- 57.
Russell, P.R. 1984. Method for expressing foreign genes in Schizosac charomyces pombe and the use in therapeutic formulations of the products, DNA constructs and transformant strains of Schizosaccharomyces pombe usable in such method and their preparation. European Patent Application EP84305571.
- 58.
Hitzeman, R.A., Chen, C.Y., Hagie, F.E., Patzer, E.J., Liu, C.-C., Estell, D.A., Miller, J.V., Yaffe, A., Kleid, D.G., Levinson, A.D. and Oppermann, H. 1983. Expression of hepatitis B virus surface antigen in yeast. Nucl. Acids. Res. 11: 2745–2763.
- 59.
Etcheverry, T., Forrester, W. and Hitzeman, R. 1986. Regulation of the chelatm promoter during the expression of human serum albumin or yeast phosphoglycerate kinase in yeast. Bio/Technology 4: 726–730.
- 60.
Fellinger, A.J., Verbakel, J.M.A., Veale, R.A., Sudbery, P.E., Bom, I.J., Overbeeke, N. and Verrips, C.T. 1991. Expression of the α-galactosidase from Cyamopsis tetragnoloba (guar) by Hansenula polymorpha. Yeast 7: 463–474.
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- Martin A. G. Gleeson
Corresponding author.
Affiliations
Salk Institute Biotechnology/Industrial Associates, Inc. (SIBIA) San Diego, CA 92037.
- Richard G. Buckholz
- & Martin A. G. Gleeson
Present address: Glaxo, Inc., 5 Moore Drive, Research Triangle Park, NC 27709.
- Richard G. Buckholz
Present address: Ligand Pharmaceuticals, Inc., 9393 Towne Centre Drive, Suite 100, San Diego, CA 92121.
- Martin A. G. Gleeson
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