Optimizing hydraulic retention time for methane production from the hydrogenic effluent left over from the co-digestion of vinasse and spent brewer's yeast cell

A rapid and selective assay was developed to measure cell surface hydrophobicity of brewer's yeast cells. During this so-called magnobead assay, bottom-fermenting yeast cells adhere to paramagnetic, polystyrene-coated latex beads which can easily be removed from the cell suspension by using a (samarium-cobalt) magnet. At pH 4 center dot 5, electrostatic repulsion between yeast cells and latex beads was found to be minimal and yeast cell adhesion was predominantly based on hydrophobic interactions. The percentage of cells adhering to the beads could be calculated and provided a measure for cell surface hydrophobicity. Cell surface hydrophobicity measured by the magnobead assay was found to yield similar results, as did determination of contact angles of water droplets on a layer of yeast cells, a standard method for measuring surface hydrophobicity. However, the magnobead assay has the following advantages: (i) it is a quick and simple method, and, more significantly, (ii) hydrophobicity can be measured under physiological conditions. Use of the magnobead assay confirmed that a higher level of cell surface hydrophobicity is correlated with stronger flocculence of brewer's lager yeast cells.

A rapid and selective assay was developed to measure cell surface hydrophobicity of brewer's yeast cells. During this so-called magnobead assay, bottom-fermenting yeast cells adhere to paramagnetic, polystyrene-coated latex beads which can easily be removed from the cell suspension by using a (samarium-cobalt) magnet. At pH 4 center dot 5, electrostatic repulsion between yeast cells and latex beads was found to be minimal and yeast cell adhesion was predominantly based on hydrophobic interactions. The percentage of cells adhering to the beads could be calculated and provided a measure for cell surface hydrophobicity. Cell surface hydrophobicity measured by the magnobead assay was found to yield similar results, as did determination of contact angles of water droplets on a layer of yeast cells, a standard method for measuring surface hydrophobicity. However, the magnobead assay has the following advantages: (i) it is a quick and simple method, and, more significantly, (ii) hydrophobicity can be measured under physiological conditions. Use of the magnobead assay confirmed that a higher level of cell surface hydrophobicity is correlated with stronger flocculence of brewer's lager yeast cells.

The highest amount of zinc (= 90%) is bound after 3 hrs of contact at low initial (total) concentrations of zinc in suspension of yeast, 10-100 mg/l at 10-30?C. The equilibrium between bound and free zinc ions is established after 6 hrs of contact time, independently on the total zinc concentration in yeast milk. No bigger changes of content of zinc bound to brewer's yeast cells was determined at temperatures 10?C and 30?C. 40% of bound zinc in the equilibrium state is bound during the first 15 min of contact of zinc ions and brewer's yeast cells at all initial (total) zinc concentrations in suspension of yeast both at 10?C and 30?C. The "KEKAM" equation can be used for the description of kinetics of zinc biosorption by waste brewer's yeast cells, for the ranges of zinc concentration 10-100 mg/l at 30?C (mean correlation coefficient 0,96) and 60,0-100 mg/l at 10?C (mean correlation coefficient 0,95).

In this study, we utilized gene knockout and overexpression techniques to generate brewer's yeast strains with either a deletion or overexpression of the POX1 gene. The strains studied included the parental strain, the POX1 deletion strain, and the POX1 overexpression strain. These strains were exposed to iso-alpha acid from hops at a concentration of 300 mg/L, leading to the induction of a viable but non-culturable (VBNC) state. Our results indicated that the silencing of the POX1 gene rendered brewer's yeast cells unable to withstand the high concentration of iso-alpha acid stress, ultimately leading to cell death. Conversely, the overexpression of POX1 accelerated the transition of yeast cells into the VBNC state compared to the parental strain. Furthermore, we evaluated the levels of reactive oxygen species (ROS), catalase (CAT) activity, superoxide dismutase (SOD) activity, glutathione reductase (GR) activity, and the mRNA expression of genes that regulate these enzymes (SYM1, CTA1, SOD1, and GLR1) in brewer's yeast cells at three distinct stages: normal, short-term stress, and VBNC states. Based on these findings, it can be inferred that the formation of the VBNC state in brewer's yeast is associated with the response to oxidative stress.

Yeast cell-agglutinating activity, designated agglutinin (possible lectin), was isolated from cell walls of both non-flocculent and flocculent brewer's yeast cells. Agglutinin-mediated aggregation of yeast cells in a manner similar to flocculation with respect to specific mannose-sensitivity, pH-dependence and calcium-dependence. Agglutinating activity was found to be heat-stable and protease-insensitive. Furthermore, addition of agglutinin to flocculent cells strongly stimulated the flocculation ability of the cells, whereas addition to non-flocculent cells rendered these cells weakly flocculent. Agglutinin was found to be released from flocculent cells during the course of a flocculation assay, but not from non-flocculent cells. Presence of mannose during the assay inhibited release of agglutinin. Our results suggest that (i) mannose-specific agglutinin is continuously synthesized during growth of brewer's yeast cells, (ii) agglutinin is present in cell walls of non-flocculent cells but is unable to bind its ligand on other cells, and (iii) the ability of yeast cells to flocculate in a flocculation assay depends, among other factors, on release of agglutinin from the cells. A 10-kDa polypeptide might represent one form of agglutinin.

A downstream process was developed for the production of yeast extract from brewer's yeast cells. Various downstream processing conditions including clarification, debittering, and the Maillard reaction were considered in the development of the process. This simple and economic clarification process used flocculating agents, specifically calcium chloride (1%). After the clarification step, a Maillard reaction is initiated as a flavor-enhancing step. By investigating the effects of several operation parameters, including the type of sugar added, sugar dosage, glycine addition, and temperature, on the degree of browning (DB), glucose addition and reaction temperature were found to have significant effects on DB. A synthetic adsorption resin (HP20) was used for the debittering process, which induced a compositional change of the hydrophobic amino acids in the yeast hydrolysate, thereby reducing the bitter taste. The overall dry matter yield and protein yield for the entire process, including the downstream process proposed for the production of brewer's yeast extract were 50 and 50%, respectively.

The purpose of this study was to assess the chromium (Cr) distribution in chromium-rich brewer's yeast cell. The chromium concentrations in the cell wall and protoplast fractions of the chromium-rich yeast were determined by neutron activation analysis (NAA). Moreover, the combined state of chromium and amino acid content in the Cr-rich brewer's yeasts was analyzed and measured. The experimental results indicate that the introduction of water-soluble chromium(III) salt as a component of the culture medium for yeasts results in a substantial amount of chromium absorbed through the cell wall by the yeast, among which 80.9% are accumulated in the protoplast. It implies that, under optimal conditions, yeasts are capable of accumulating large amounts of chromium and incorporating chromium into organic compounds.

The nucleotide sequence of a 1.4-kilobase DNA fragment containing the alpha-acetolactate decarboxylase gene of Enterobacter aerogenes was determined. The sequence contains an entire protein-coding region of 780 nucleotides which encodes an alpha-acetolactate decarboxylase of 260 amino acids. The DNA sequence coding for alpha-acetolactate decarboxylase was placed under the control of the alcohol dehydrogenase I promoter of the yeast Saccharomyces cerevisiae in a plasmid capable of autonomous replication in both S. cerevisiae and Escherichia coli. Brewer's yeast cells transformed by this plasmid showed alpha-acetolactate decarboxylase activity and were used in laboratory-scale fermentation experiments. These experiments revealed that the diacetyl concentration in wort fermented by the plasmid-containing yeast strain was significantly lower than that in wort fermented by the parental strain. These results indicated that the alpha-acetolactate decarboxylase activity produced by brewer's yeast cells degraded alpha-acetolactate and that this degradation caused a decrease in diacetyl production.

A new approach for the immobilization of permeabilized brewer's yeast cells in a modified composite polyvinyl alcohol lens-shaped capsule containing montmorillonite and dimethyldioctadecylammonium bromide for use as a biocatalyst

Brewer's yeast cell wall (BYC) is a valuable foodstuff that prevents constipation in rats. In this study, the effects of yogurt supplemented with brewer's yeast cell wall (BYC-Y) on constipated male Sprague-Dawley rats induced by loperamide (2 mg/kg body weight) were investigated. The preventive effect of BYC-Y on experimentally constipated rats was examined and compared with that of nonfiber diet (Control) and standard yogurt (Yogurt) as the control diets. The number of feces and fecal dry weight were greater in rats fed with BYC-Y than in those fed with Control or Yogurt. Moreover, the effect of BYC on the improvement of cecal microflora in constipated experimental model rats was evaluated. The number of Lactobacillus, Eubacterium, and total anaerobes per unit cecal contents increased (p<0.03) as a result of BYC ingestion. These findings indicate that BYC-Y or BYC is effective for the alleviation of constipation and bowel environment.

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).

Drying enhances immunoactivity of spent brewer's yeast cell wall β-d-glucans

Effects of treatments with proteolytic enzymes and protein-modifying reagents on flocculation of brewer's yeast IFO 2018 were investigated. The floc-forming ability of the yeast cells was irreversibly eliminated by treatment with papain, trypsin, chymotrypsin or pepsin, indicating that certain proteins on the cell surface participate in the yeast flocculation. Chemical modification with reagents, known to act on disulfide bridges, carboxyl and/or phosphate groups, phenolic groups, amino groups, and imidazole groups, also destroyed the ability to flocculate, although in some cases a high concentration (8 M) of urea was necessary in addition to protein-modifying reagents. Thus, it is suggested strongly that these functional groups of amino acid residues of the proteins are essential for the floc-forming ability of brewer's yeast cells.

This study was designed to determine whether yogurt supplemented with insoluble dietary fibers would lower the levels of serum lipids in 89 normal and hyper-cholesterolemic adults. Yogurt supplemented with brewer’s yeast cell wall (BYC) was administered to 89 volunteers, and its effects on serum cholesterol levels were examined. The yoturt used contained 6g of BYC per 200ml, and an other batch without BYC served as control diet. The volunteers ingested 200ml of BYC-yogurt (BYC-Y) or control yogurt (CT-Y) everyday for four weeks in a crossover experiment. Blood samples were drawn from the subjects for analysis on five occasions two weeks before the start of the experimental period, and immediately before and after each dietary period. It was observed that the levels of the serum total cholesterol (TC), high-density lipoprotein (HDL) cholesterol and low-density lipoprotein (LDL) cholesterol significantly decreased following ingestion of BYC-Y compared with those before BYC-Y ingestion. Furthermore, the TC levels of the subjects (serum TC≥200mg/dl) taking the BYC-Y diet were found to be significantly lower than those taking the CT-Y diet. However, the HDL- and LDL-chloesterol levels of those taking the BYC-Y diet were not found to be significantly different from those taking the CT-Y diet. No abnormalities in serum biochemical parameters were observed during the experimental period in all the subjects. These results indicate that taking BYC-Y is effective in lowering the serum cholesterol levels in subjects with the tendency for hypercholesterolemia. The results suggest that BYC-Y may be utilized as a food for controlling hypercholesterolemia.

Brewer's yeast cell wall (BYC) was tested on constipated male Sprague-Dawley rats that had been induced by loperamide (2 mg/kg of body weight). The preventive effect of BYC on constipation was examined and compared with that of a non-fiber diet (NF) as the control. The dose-response of BYC and the effect on defecation by constipated experimental rats were also compared with the characteristics of cellulose diet (CE) group which served as a control. Defecation was observed to be greater by the rats fed with BYC than by those fed with NF or CE. The fecal water content and level of volatile fatty acids (VFA) in the cecal contents were likewise higher in the rats fed with BYC. These results indicate that the administration of BYC was effective for improving defecation and other parameters related to defecation. These favorable effects of BYC supplemented to the diet are attributed to the fermentation ability, water holding capacity and swelling force in the large intestine.