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M Stratford

Publications and source records attributed to M Stratford.

10 recordsLinked to original sources
PubMed1992

Yeast flocculation: reconciliation of physiological and genetic viewpoints.

Yeast flocculation results from surface expression of specific proteins (lectins). Two flocculation phenotypes were suggested by physiological and biochemical tests, whereas genetic data suggested a larger number of mechanisms of flocculation. After reviewing the biochemistry, physiology and genetics of flocculation, a new hypothesis combining the data available from these different sources, is proposed. Flocculation results when lectins present on flocculent cell walls bind to sugar residues of neighbouring cell walls. These sugar receptors are intrinsic to the mannan comprising cell walls of Saccharomyces cerevisiae. Two lectin phenotypes were revealed by sugar inhibition studies. The gluco- and mannospecific NewFlo phenotype is not, as yet, found in genetically defined strains. Mannospecific flocculation (Flo1 phenotype) is found in strains containing the genes FLO1, FLO5 and FLO8. This phenotype is also found following mutation of the TUP1 or CYC8 loci, in previously non-flocculent strains. It is therefore proposed that the structural gene for mannospecific flocculation is common or possibly ubiquitous in non-flocculent strains and in consequence, FLO1, FLO5 and FLO8 are probably regulatory genes, exerting positive control over the structural gene. Flocculation expression requires lectin secretion to the cell surface. Many of the observed 'suppressions' of flocculation may be due to mutations of the secretory process, involved in transporting structural proteins to the cell wall. The possible involvement of killer L double-stranded RNA with flocculation is suggested, given the lectin properties of viral coat proteins and an association between L double-stranded RNA and the Flo1 phenotype.

Cell Aggregation↗
PubMed1992

Yeast flocculation: receptor definition by mnn mutants and concanavalin A.

Yeast flocculation involves the binding of surface lectins on flocculent yeasts, to carbohydrate receptors present as constituents of yeast cell walls. Receptors were investigated by coflocculation of flocculent strains of Saccharomyces cerevisiae, of both Flo 1 and NewFlo phenotypes, to known mnn mutants which vary in the wall mannan structure. Strong coflocculation was found with mnn1, mnn4, mnn9 and control strains, while very little coflocculation was found with mnn2 and mnn5 strains. In contrast, aggregation of these mutants by concanavalin A, a lectin with similar sugar inhibition to NewFlo phenotype flocculation, showed strong aggregation of mnn1, mnn4 and mnn5 strains and poor aggregation of mnn2 and mnn9 strains. The mmn mutant data suggested that flocculation receptors were the outer-chain mannan side-branches, two or three mannose residues in length, confirming an earlier theory based on sugar inhibition data. The similarities and differences between flocculation and concanavalin A aggregation are discussed.

Carbohydrate Sequence↗
PubMed1989

Evidence for two mechanisms of flocculation in Saccharomyces cerevisiae.

Inhibition of flocculation by sugars was studied in 41 strains of Saccharomyces cerevisiae. Two distinct groupings were found. The first type, including all strains containing FLO 1, FLO 4, FLO 5, FLO 8 and TUP 1, were partially inhibited by mannose only. The second type (NewFLO) were completely inhibited by mannose, maltose, glucose and sucrose. Sugar inhibitions were reversible and were exacerbated by increased agitation. NewFLO strains were more sensitive to inhibition by inorganic salts than the FLO 1 type strains. This inhibition is probably chaotropic in nature. Most FLO 1 type strains are constitutive, expressing flocculation throughout growth, whereas, NewFLO strains are completely repressed by the presence of excess ammonium ions. It is proposed that this indicates two entirely distinct mechanisms of flocculation, probably involving a mannose-specific lectin in the FLO 1 type and a broad specificity lectin in the NewFLO type.

Calcium Chloride↗
PubMed1988

Yeast flocculation: quantification.

Yeast flocculation is an orthokinetic process dependent upon mechanical agitation for all quantitative measurements. From several methods which were assessed, orbital shaking was selected as being the most practical as well as producing the most meaningful results. Quantitative measurements of flocculation were made in terms of minimum agitation threshold, initial rate, extent of flocculation at equilibrium and flocculated particle size at equilibrium. All these parameters were strain dependent. Critical cell density functions were formed if agitation was limiting regardless of how the agitation was imposed, and are unlikely to be related to bond strength.

Colony Count, Microbial↗
PubMed1988

Yeast flocculation: a dynamic equilibrium.

The steady state in yeast flocculation is a dynamic equilibrium between flocculated and dispersed yeast cells. The free cell concentration is directly proportional to the total cell concentration and may be expressed as an equilibrium constant. Increased agitation decreases floc size and equilibrium constant whilst increasing floc-surface area and free-cell concentration. Values of equilibrium constant are influenced by agitation in a complex relationship probably involving the floc-surface area and floc momentum. Inhibition of flocculation by mannose and low pH is reversible and becomes greater with increased agitation. Both these inhibitions appear consistent with a weakening of flocculent bond strength by these inhibitors.

Colony Count, Microbial↗
PubMed1987

Yeast flocculation: kinetics and collision theory.

Flocculent yeast cells have an absolute requirement for mechanical energy input in order for flocculation to occur. Flocculation is arrested by cessation of energy input. The initial rate of flocculation increases as the square of the cell concentration. There is a minimum shaking speed to initiate flocculation and thereafter the initial rate of flocculation increases exponentially with the shaking speed. The minimum shaking speed for flocculation to occur increases with pH value. Activation energy for flocculation, derived from Arrhenius-like plots, varies with pH value. We propose that activation energy is required to overcome mutual repulsion between charged yeast cells and allow flocculent bonds to be formed.

Calcium↗
PubMedDate not supplied

Yeast flocculation: Flo1 and NewFlo phenotypes and receptor structure.

Flocculation characteristics of 42 flocculent strains of Saccharomyces cerevisiae were examined. Two entirely distinct 'lectin-like' mechanisms of flocculation were distinguished by sugar, salt, and low pH inhibitions, protease sensitivity, and selective expression of flocculation. One group, termed Flo1 phenotype, was inhibited by mannopyranoses and contained all strains bearing known genes affecting flocculation. The other group, termed NewFlo phenotype, contained the majority of brewery ale stains and was inhibited by manno- and glucopyranoses. Detailed sugar-inhibition work revealed the probable receptor identity of both Flo1 and NewFlo flocculation, as being non-reducing termini of alpha-(1-3)-linked mannan side branches, two or three mannopyranose residues in length.

Carbohydrates↗