Research Paper | Published:

Saccharomyces Cerevisiae Strains that Overexpress Heterologous Proteins

Bio/Technologyvolume 9pages183187 (1991) | Download Citation



We describe a system that facilitates the selection of host mutants that overproduce a range of secreted and internally produced heterologous proteins in Saccharomyces cerevisiae. These mutants were initially selected for their ability to oversecrete recombinant human albumin (rHA), as detected by a direct visual assay that relies upon antibody precipitation in solid media. Yeast strains that were able to synthesize and secrete increased levels of rHA also produced elevated levels of internally expressed proteins including α1-antitrypsin Pittsburgh variant and plasminogen activator inhibitor type 2.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1

    Aharonowitz, Y. and Cohen, G. 1981. The microbiological production of pharmaceuticals. Sci. Amer. 245: 106–119.

  2. 2

    Loison, G., Videl, A., Findeli, A., Roitsch, C., Balloul, J.M. and Lemoine, Y. 1989. High level of expression of a protective antigen of schistosomes in Saccharomyces cerevisiae. Yeast 5: 497–507.

  3. 3

    Hallewell, R.A., Mills, R., Tekamp-Olson, P., Blacker, R., Rosenberg, S., Oetting, F., Masiarz, F.R. and Scandella, C.J. 1987. Amino terminal acetylation of authentic human Cu, Zn superoxide dismutase produced in yeast. Bio/Technology 5: 363–366.

  4. 4

    Hiramatsu, R., Aikaura, J., Horinouchi, S. and Beppu, T. 1989. Secretion by yeast of the zymogen form of Mucor rennin, an aspartic proteinase of Mucor pusillus, and its conversion to the mature form. J. Biol. Chem. 262: 16862–16865.

  5. 5

    Kriskern, P.J., Hagopian, A., Montgomery, D.L., Burke, P., Dunn, N.R., Hofmann, K.J., Miller, W.J. and Ellis, R.W. 1986. Unusually high-level expression of a foreign gene (hepatitis B core antigen) in Saccharomyces cerevisiae. Gene 46: 135–141.

  6. 6

    Cousens, L.S., Shuster, J.R., Gallegos, C., Stempien, M.M., Urdea, M.S., Sanchez-Pescador, R., Taylpr, A. and Tekamp-Olson, P. 1987. High-level expression of proinsulin in the yeast Saccharomyces cerevisiae. Gene 61: 265–275.

  7. 7

    Ecker, D.J., Stadel, J.M., Butt, T.R., March, J.A., Monia, B.P., Powers, D.A., German, J.A., Clark, P.E., Warren, F., Shatzman, A. and Crooke, S.T. 1989. Increasing gene expression in yeast by fusion to ubiquitin. J. Biol. Chem. 264: 7715–7719.

  8. 8

    Sabin, E.A., Lee-Hu, C.T., Shuster, J.R. and Barr, P.J. 1989. High-level expression and in vivo processing of chimeric ubiquitin fusion proteins in Saccharomyces cerevisiae. Bio/Technology 7: 705–709.

  9. 9

    Kaisho, Y., Yoshimura, K. and Hakahama, K. 1989. Increase in gene expression by respiratory-deficient mutant. Yeast 5: 91–98.

  10. 10

    Smith, R.A., Duncan, M.J. and Moir, D.J. 1985. Heterologous protein secretion from yeast. Science 229: 1219–1224.

  11. 11

    Rudolph, H.K., Antebi, A., Fink, G.R., Buckley, C.M., Dorman, T.E., Le Vitre, J., Davidson, L.S., Mao, J. and Moir, D.T. 1989. The yeast secretory pathway is perturbed by mutations in PMR1, a member of a Ca2+ ATPase family. Cell 58: 133–145.

  12. 12

    Sakai, A., Shimiza, Y. and Hishinama, F. 1988. Isolation and characterisation of mutants which show an oversecretion phenotype in Saccharomyces cerevisiae. Genetics 119: 499–506.

  13. 13

    Suzuki, K., Ichikawa, K. and Yoshifumi, J. 1989. Yeast mutants with enhanced ability to secrete human lysozyme: isolation and identification of a protease-deficient mutant. Mol. Gen. Genet. 219: 58–64.

  14. 14

    Moehle, C.M., Tizard, R., Lemmon, S.K., Smart, J. and Jones, E.W. 1987. Protease B of the lysozyme-like vacuole of the yeast Saccharomyces cerevisiae is homologous to the subtilisn family of serine protease. Mol. Cell. Biol. 7: 4390–4399.

  15. 15

    Moehle, C.M., Aynardi, M.W., Kolodny, M.R., Park, F.J. and Jones, E.W. 1987. Protease B of Saccharomyces cerevisiae: Isolation and regulation of the PRB1 structural gene. Genetics 115: 255–263.

  16. 16

    Moehle, C.M. and Jones, E.W. 1990. Consequences of growth media, gene copy number and regulatory mutations on the expression of the PRB1 gene of Saccharomyces cerevisiae. Genetics 124: 39–55.

  17. 17

    Saheki, T. and Holzer, H. 1975. Proteolytic activities in yeast. Biochem. Biophys. Acta. 384: 203–214.

  18. 18

    Wolf, D. and Ehmann, C. 1979. Studies on a protease B mutant of yeast. Eur. J. Biochem. 98: 375–384.

  19. 19

    Zubenko, G. and Jones, E. 1981. Protein degradation, meiosis and sporulation in proteinase-deficient mutants of Saccharomyces cerevisiae. Genetics 97: 45–64.

  20. 20

    Yanish-Perron, C., Vieira, J. and Messing, J. 1985. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC9 vectors. Gene 33: 103–119.

  21. 21

    Goodey, A.R., Doel, S.M., Piggott, J.R., Watson, M.E. and Carter, B.L.A. 1987. Expression and secretion of foreign polypeptides in yeast, p. 401–429. In: Yeast Biotechnology. Berry, D.R., Russell, I. and Stewart, G.G. (Eds.). Allen Unwin, UK.

  22. 22

    Sleep, D., Belfield, G.P. and Goodey, A.R. 1990. The secretion of human serum albumin from the yeast Saccharomyces cerevisiae using five different leader sequences. Bio/Technology 8: 42–46.

  23. 23

    Chinery, S.A. and Hinchliffe, E. 1989. A novel class of vector for yeast transformation. Curr. Genet. 16: 21–25.

  24. 24

    Andreadis, A., Hsu, Y.-P., Hermodson, M., Kohlaw, G. and Shimmel, P. 1984. Yeast LEU2. Jour. of Biol. Chem. 259: 8059–8062.

  25. 25

    Jones, E.W. 1977. Proteinase mutants of Saccharomyces cerevisiae. Genetics 18: 233–270.

  26. 26

    Sleep, D., Ogden, J.E., Roberts, N.A. and Goodey, A.R. 1990. Cloning and characterisation of the Saccharomyces cerevisiae glycerol-3-phosphate dehydrogenase promoter. Gene. In press.

  27. 27

    Steven, J., Cottingham, I.R., Berry, S.J., Goodey, A.R., Courtney, M.G. and Ballance, D.J. 1990. Expression of plasminogen activator inhibitor 2 in Saccharomyces cerevisiae. Submitted.

  28. 28

    Hannahan, D. 1983. Studies on transformation of Escherichia coli with plasmids. J. Mol. Biol. 166: 557–580.

  29. 29

    Mandel, M. and Higa, A. 1970. Calcium dependent bacteriophage DNA infection. J. Mol. Biol. 53: 159–162.

  30. 30

    Beggs, J.D. 1978. Transformation of yeast by a replicating hybrid plasmid. Nature 275: 104–109.

  31. 31

    Vogelstein, B. 1987. Rapid purification of DNA from agarose gels by centrifugation through a disposable plastic column. Anal. Biochem. 160: 115–118.

  32. 32

    Birnboim, H.C. and Doly, J. 1979. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 7: 1513–1523.

  33. 33

    Linquist, S. 1981. Regulation of protein synthesis during heat shock. Nature 293: 311–314.

  34. 34

    Thomas, P.S. 1980. Hybridisation of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc. Natl. Acad. Sci. USA 77: 5201–5205.

  35. 35

    Tourbin, H., Staehelin, T. and Gordon, J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76: 4350–4354.

  36. 36

    Owen, M.C., Brennan, S.O., Lewis, J.H. and Carrell, R.W. 1983. Mutation of antitrypsin to antithrombin α1-antitrypsin Pittsburgh (358 Met→Arg) a fatal bleeding disorder. N. Engl. J. Med. 309: 694–690.

  37. 37

    Futcher, B. 1988. The 2μm circle plasmid of Saccharomyces cerevisiae. Yeast 4: 27–40.

  38. 38

    Hinchliffe, E., Kenny, E. and Leaker, A. 1987. Novel products from surplus yeast via recombinant DNA technology. European Brewery Convention Symposium on Brewing Yeast, Monogr. XIII: 139–154.

  39. 39

    Hoekema, A., Kastelein, R.A., Vasser, M. and de Boer, H. A. 1987. Codon replacement in the PGK1 gene of Saccharomyces cerevisiae: Experimental approach to study the role of biased codon usage in gene expression. Mol. and Cell Biol. 7: 2914–2924.

Download references

Author information

Author notes

  1. D. Sleep: Corresponding author.


  1. Delta Biotechnology Limited, Castle Court, 59 Castle Boulevard, Nottingham, NG7 1FD, Great Britain

    • D. Sleep
    • , G.P. Belfield
    • , D.J. Ballance
    • , J. Steven
    • , S. Jones
    • , L.R. Evans
    • , P.D. Moir
    •  & A.R. Goodey


  1. Search for D. Sleep in:

  2. Search for G.P. Belfield in:

  3. Search for D.J. Ballance in:

  4. Search for J. Steven in:

  5. Search for S. Jones in:

  6. Search for L.R. Evans in:

  7. Search for P.D. Moir in:

  8. Search for A.R. Goodey in:

About this article

Publication history



Issue Date


Further reading