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. 1994 Aug;60(8):2754–2758. doi: 10.1128/aem.60.8.2754-2758.1994

Purification and partial characterization of a flocculin from brewer's yeast.

M H Straver 1, G Smit 1, J W Kijne 1
PMCID: PMC201719  PMID: 8085818

Abstract

Analysis of a shear supernatant from flocculent, "fimbriated" Saccharomyces cerevisiae brewer's yeast cells revealed the presence of a protein involved in flocculation of the yeast cells and therefore designated a flocculin. The molecular mass of the flocculin was estimated to be over 300 kDa, as judged from sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gel permeation chromatography of the flocculin yielded an aggregate with an apparent molecular weight of > 2,000. The flocculin was found to be protease sensitive, and the sequence of its 16 N-terminal amino acids revealed at least 69% identity with the predicted N terminus of the putative protein encoded by the flocculation gene FLO1. The flocculin was isolated from flocculent S. cerevisiae cells, whereas only a low amount of flocculin, if any, could be isolated from nonflocculent cells. The flocculin was found to stimulate the flocculation ability of flocculent yeast cells without displaying lectinlike activity (that is, the ability to agglutinate yeast cells).

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Selected References

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  1. Hewick R. M., Hunkapiller M. W., Hood L. E., Dreyer W. J. A gas-liquid solid phase peptide and protein sequenator. J Biol Chem. 1981 Aug 10;256(15):7990–7997. [PubMed] [Google Scholar]
  2. Kihn J. C., Masy C. L., Mestdagh M. M., Rouxhet P. G. Yeast flocculation: factors affecting the measurement of flocculence. Can J Microbiol. 1988 Jun;34(6):779–781. doi: 10.1139/m88-132. [DOI] [PubMed] [Google Scholar]
  3. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [PubMed] [Google Scholar]
  4. Miki B. L., Poon N. H., James A. P., Seligy V. L. Possible mechanism for flocculation interactions governed by gene FLO1 in Saccharomyces cerevisiae. J Bacteriol. 1982 May;150(2):878–889. doi: 10.1128/jb.150.2.878-889.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Schuster R. Determination of amino acids in biological, pharmaceutical, plant and food samples by automated precolumn derivatization and high-performance liquid chromatography. J Chromatogr. 1988 Oct 14;431(2):271–284. doi: 10.1016/s0378-4347(00)83096-0. [DOI] [PubMed] [Google Scholar]
  6. Smit G., Straver M. H., Lugtenberg B. J., Kijne J. W. Flocculence of Saccharomyces cerevisiae cells is induced by nutrient limitation, with cell surface hydrophobicity as a major determinant. Appl Environ Microbiol. 1992 Nov;58(11):3709–3714. doi: 10.1128/aem.58.11.3709-3714.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Stratford M., Carter A. T. Yeast flocculation: lectin synthesis and activation. Yeast. 1993 Apr;9(4):371–378. doi: 10.1002/yea.320090407. [DOI] [PubMed] [Google Scholar]
  8. Stratford M. Yeast flocculation: flocculation onset and receptor availability. Yeast. 1993 Jan;9(1):85–94. doi: 10.1002/yea.320090111. [DOI] [PubMed] [Google Scholar]
  9. Stratford M. Yeast flocculation: receptor definition by mnn mutants and concanavalin A. Yeast. 1992 Aug;8(8):635–645. doi: 10.1002/yea.320080807. [DOI] [PubMed] [Google Scholar]
  10. Stratford M. Yeast flocculation: reconciliation of physiological and genetic viewpoints. Yeast. 1992 Jan;8(1):25–38. doi: 10.1002/yea.320080103. [DOI] [PubMed] [Google Scholar]
  11. Straver M. H., Kijne J. W., Smit G. Cause and control of flocculation in yeast. Trends Biotechnol. 1993 Jun;11(6):228–232. doi: 10.1016/0167-7799(93)90133-T. [DOI] [PubMed] [Google Scholar]
  12. Straver M. H., vd Aar P. C., Smit G., Kijne J. W. Determinants of flocculence of brewer's yeast during fermentation in wort. Yeast. 1993 May;9(5):527–532. doi: 10.1002/yea.320090509. [DOI] [PubMed] [Google Scholar]
  13. Teunissen A. W., Holub E., van der Hucht J., van den Berg J. A., Steensma H. Y. Sequence of the open reading frame of the FLO1 gene from Saccharomyces cerevisiae. Yeast. 1993 Apr;9(4):423–427. doi: 10.1002/yea.320090413. [DOI] [PubMed] [Google Scholar]
  14. Watari J., Takata Y., Ogawa M., Sahara H., Koshino S., Onnela M. L., Airaksinen U., Jaatinen R., Penttilä M., Keränen S. Molecular cloning and analysis of the yeast flocculation gene FLO1. Yeast. 1994 Feb;10(2):211–225. doi: 10.1002/yea.320100208. [DOI] [PubMed] [Google Scholar]
  15. van Loosdrecht M. C., Lyklema J., Norde W., Schraa G., Zehnder A. J. Electrophoretic mobility and hydrophobicity as a measured to predict the initial steps of bacterial adhesion. Appl Environ Microbiol. 1987 Aug;53(8):1898–1901. doi: 10.1128/aem.53.8.1898-1901.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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