Isolation and characterization of a polypeptide absent from non-flocculent mutants of Saccharomyces cerevisiae

Holmberg, Steen

doi:10.1007/bf02906111

Cited by 34 publications

(16 citation statements)

References 21 publications

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“…Treatment of flocculent cells with proteinases, protein-denaturing and protein-modifying reagents (Nishihara et al, 1977(Nishihara et al, , 1982 has indicated an important role for protein components of the cell wall in the mechanism of floc formation. Polypeptides specific to cell-wall extracts from flocculent strains have been reported (Holmberg, 1978).…”

Section: Introductionmentioning

confidence: 99%

Discrimination by Heat and Proteinase Treatments between Flocculent Phenotypes Conferred on Saccharomyces cerevisiae by the Genes FLO1 and FLO5

1985

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The effects of elevated temperature and of digestion with a variety of proteinases on the flocforming ability of flocculent strains of Saccharomyces cerevisiae, both genetically defined (FLO1 and FLO5) laboratory and genetically undefined brewing strains, have been determined. This has permitted classification of the flocculent phenotypes of these strains according to criteria other than quantitative grading of flocculence. The flocculent phenotypes conferred by both the FLO1 and the FLO5 gene were irreversibly lost upon treatment with pronase, proteinase K, trypsin or 2-mercaptoethanol treatments. However, the floc-forming ability of cells of the FLO1 strain ABXL-1D was destroyed by chymotrypsin digestion and was stable to incubation at 70 degrees C, whereas the floc-forming ability of cells of the FLO5 strain ABXR-11A was resistant to the action of chymotrypsin and was heat labile. Tetrad analysis of a cross of these FLO1 and FLO5 strains indicated that the chymotrypsin and heat sensitivity phenotypes were FLO-gene determined. It appears that expression of the FLO1 and FLO5 genes leads to the production of different and characteristic cell-wall proteins underlying their respective flocculent phenotypes.

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Section: Introductionmentioning

confidence: 99%

Discrimination by Heat and Proteinase Treatments between Flocculent Phenotypes Conferred on Saccharomyces cerevisiae by the Genes FLO1 and FLO5

1985

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“…The presence of a 37 kDa protein in extracts of flocculent cultures was reported for S. cerevisiae strains [20], K. marxianus strains [11,21,22], and flocculent Hansenula anomala strains [23]. A polypeptide of 13 kDa was also identified in alkaline cell extracts of flocculent S. cerevisae [24,25], as well as a 25 kDa protein [26]. In the K. marxianus case, the involvement of p37 in flocculation was recently supported by the fact that a K. marxianus mutant, deficient in the synthesis of p37, was no longer able to flocculate [27].…”

Section: Basics Of Yeast Flocculation Yeast Flocculation Mechanismmentioning

confidence: 82%

Applications of yeast flocculation in biotechnological processes

Domingues

Vicente

Lima

et al. 2000

Biotechnol. Bioprocess Eng.

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A review on the main aspects associated with yeast flocculation and its application in biotechnological processes is presented. This subject is addressed following three main aspects -the basics of yeast flocculation, the development of "new" flocculating yeast strains and bioreactor development. In what concerns the basics of yeast flocculation, the state of the art on the most relevant aspects of mechanism, physiology and genetics of yeast flocculation is reported. The construction of flocculating yeast strains includes not only the recombinant constitutive flocculent brewer's yeast, but also recombinant flocculent yeast for lactose metabolisation and ethanol production. Furthermore, recent work on the heterologous β-galactosidase production using a recombinant flocculent Saccharomyces cerevisiae is considered. As bioreactors using flocculating yeast cells have particular properties, mainly associated with a high solid phase hold-up, a section dedicated to its operation is presented. Aspects such as bioreactor productivity and culture stability as well as bioreactor hydrodynamics and mass transfer properties of flocculating cell cultures are considered. Finally, the paper concludes describing some of the applications of high cell density flocculation bioreactors and discussing potential new uses of these systems.

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“…The amino acid components of 37 Kprotein were abundant in Asx, Thr, Phe, and Tyr. Holmberg 17 ) also isolated a 13K dalton peptide present in the alkaline extract from a flocculent strain of S·. cerevisiae.…”

Section: Discussionmentioning

confidence: 99%

Flocculation Mechanism ofHansenula anomalaJ224

Saito¹,

Sato²,

Shimoi³

et al. 1990

Agricultural and Biological Chemistry

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The flocculation mechanism of a flocculent yeast, Hansenula anomalaJ224, isolated from a waste water treatment tank of a Japanese sake brewing factory, was investigated by comparing some characteristics with strain J224-1 (a nonflocculent mutant of strain J224). The cell titration curves showed that strain J224 was different from strain J224-1 in charge distribution on the cell surface. The cell wall of strain J224· was more abundant in Mg 2 +, Ca 2 +, Na +, NH 4 +, and total phosphorus contents but had less fatty acids than that of strain J224-1. The cell walls of both strains had almost the same amino acid composition. Acylation of strain J224 with acetic anhydride caused deflocculation. By treatment with proteinase K, an additional protein of 37 K dalton (37 K-protein) whose molecular mass was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was released in the supernatant from strain J224 cells. The 37 K-protein was purified by cation-exchange highperformance liquid chromatography. The amino acid composition of this protein was different from that of the cell wall. Flocculation of strain J224 was promoted by the addition of 37 K-protein.We discussed the involvement of ionic interaction between 37 K-protein and phosphate and hydrophobic interaction between 37 K-proteins on the cell surface of adjacent cells in the flocculation of strain J 224. 1425For waste water treatments, we have developed yeasts to assimilate raw starch effectivelyand maintain cell number in a waste water treatment tank under inefficient nutritional conditions, and isolated a flocculent yeast, Hansenula anomala J224, from a waste water treatment tank of a Japanese sake brewing factory. We reported the ability of this strain to remove waste loads in various waste waters, its flocculent characteristics, and the merits of using this strain previously.1,2) F or the mechanism of flocculation of yeasts, there are many studies on brewer's yeast (Saccharomyces cerevisiae) and genes encoded the floc-forming ability have also been studied.3 )It was generally recognized that flocculation was a specific cell adhesion, with proteinaceous, lectin-like molecules bound to amannan carbohydrates of adjoining cells and that bivalent metal ions, in particular Ca 2 +, were very important in this mechanism. -6 )Unlike brewer's yeasts, strain J224 does not need bivalent metal ions for flocculation. The floc-forming ability of strain J224 is not inhibited by the addition of various sugars (mannose, glucose, a-methyl-mannoside etc.) and is only eliminated irreversibly by treatment with limited kinds of proteolytic enzymes, pronase E or proteinase K. The comparison of characteristics between flocculent and nonflocculent cells is an useful tool to elucidate the mechanism of flocculation. Then we obtained a nonflocculent mutant, J224-1, from the flocculent strain J224.2 ) In addition, pronase E or proteinase K-treated strain J224 was reduced in hydrophobicity to the level of strain J224-1 with elimination of the floc-forming ability.2)In this p...

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Isolation and characterization of a polypeptide absent from non-flocculent mutants of Saccharomyces cerevisiae

Cited by 34 publications

References 21 publications

Discrimination by Heat and Proteinase Treatments between Flocculent Phenotypes Conferred on Saccharomyces cerevisiae by the Genes FLO1 and FLO5

Discrimination by Heat and Proteinase Treatments between Flocculent Phenotypes Conferred on Saccharomyces cerevisiae by the Genes FLO1 and FLO5

Applications of yeast flocculation in biotechnological processes

Flocculation Mechanism ofHansenula anomalaJ224

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