A rapid and selective assay for measuring cell surface hydrophobicity of brewer's 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 4·5, 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.     

Isolation and partial purification of mannose-specific agglutinin from brewer's yeast involved in flocculation

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.     

Effects of soy protein hydrolysates on the growth and fermentation performances of brewer's yeast

Soy protein isolate was hydrolysed with Alcalase, Papain, Flavorzyme and Protemax, respectively, and further fractioned by ultrafiltration. The resulting soy protein hydrolysates (SPH) and their ultrafiltration fractions were used to examine their effects on the growth and fermentation performances of brewer's yeast. Results showed that degree of hydrolysis, molecular weight distribution and amino acid composition of SPH significantly affected the growth, viability and fermentation performance of brewer's yeast. The SPH prepared from different proteolytic enzymes exhibited distinct growth‐ and fermentation‐promoting activity for brewer's yeast. The SPH treated with Protemax for 9 h and with the molecular weight below 3 kDa showed the highest growth‐promoting activity and induced more rapidly reducing sugar consumption and higher ethanol production. The relatively lower molecular weight and the hydrophilic and electropositive amino acid residues (Lys, His, Arg and Ile) in SPH might be responsible for its functionality, promoting the growth and fermentation of brewer's yeast.     

Effects of wheat gluten hydrolysates fractionated by different methods on the growth and fermentation performances of brewer's yeast under high gravity fermentation

Wheat gluten hydrolysates (WGH) were fractionated through ultrafiltration membrane with molecular weight (Mw) cut‐off of 3 kDa and ethanol precipitation, respectively. WGH and their fractions were used to examine their effects on the growth and fermentation performances of brewer's yeast under high gravity fermentation. Results showed that WGH and their fractions exhibited significant differences in biomass accumulation, viability, ethanol yield, free amino nitrogen and sugar consumptions under high gravity fermentation. Compared to WGH, the fractions with Mw < 3 kDa and the supernatant of WGH treated with ethanol precipitation showed better fermentation performance for brewer's yeast. The relatively lower molecular weight and the higher levels of Leu, Lys, His and Arg in these two fractions might be responsible for their bioactivity for brewer's yeast. Thus, both ultrafiltration and ethanol precipitation could be used as efficient methods for enriching peptides with significant growth‐ and fermentation‐promoting activity for brewer's yeast under high gravity fermentation.     

外源海藻糖对啤酒酵母在热胁迫下的保护作用

为探究外源海藻糖对啤酒酵母在热胁迫下的保护作用,作者在3个热胁迫条件下检测外源海藻糖对啤酒酵母活力及活性的影响,并通过分析转录组数据对热胁迫下外源海藻糖对啤酒酵母的保护作用机理进行初步分析。结果表明,外源海藻糖可以提升啤酒酵母的耐热性,但具有一定限度,温度的升高和胁迫时间的延长导致不同浓度海藻糖的作用差别减小。另外,添加海藻糖可以增加酵母胞内海藻糖质量分数,添加海藻糖对啤酒酵母转录水平上的影响主要体现在核糖体的合成和相关功能上。结合本研究推测,外源海藻糖对啤酒酵母在热胁迫下的保护作用是通过增加胞内海藻糖质量分数实现的,上升的胞内海藻糖质量分数促进了核糖体合成及相关代谢,使核糖体功能增强,进而实现对啤酒酵母耐热性的增强。     

Use of β-glucan from spent brewer's yeast as a thickener in skimmed yogurt: Physicochemical,textural, and structural properties related to sensory perception

Powdered β-glucan extracted from brewer's yeast (Yestimun, Leiber GmbH, Bramsche, Germany) was incorporated into skimmed-milk yogurt at varying concentrations (0.2–0.8% wt/wt) to investigate its potential application as a thickener. The effect of β-glucan fortification on the nutritional profile, microstructure, physicochemical properties, and texture of freshly prepared yogurts was investigated. Sensory evaluation was also conducted and was correlated with instrumental analysis. The addition of Yestimun significantly reduced the fermentation time of the yogurt mix from 4 h to 3 h. Scanning electron microscopy revealed that β-glucan particles formed small spherical clusters within the yogurt matrix. The majority of the physicochemical properties (syneresis, viscosity, color, and titratable acidity) remained unaffected by the incorporation of Yestimun in the recipe. Textural properties showed a gradual increment with increasing β-glucan concentration. Hardness, total work done, adhesive force, and adhesiveness increased by 19.27, 23.3, 21.53, and 20.76%, respectively, when using the highest amount of Yestimun powder. Sensory analysis (n = 40) indicated that fortifying yogurt with Yestimun at 0.8% (wt/wt) concentration may affect overall acceptance ratings, which was attributed to adverse flavor and aftertaste effects. However, the overall liking score of the yogurt (5.0/9.0) shows potential for commercialization of the product.     

酿酒废酵母吸附共存重金属离子的研究

为研究酿酒废酵母吸附共存重金属离子的可行性,在单因素试验的基础上,采用响应曲面法探讨酿酒废酵母吸附共存离子Pb2+、Ni 2+、Cu2+的最佳工艺条件。结果表明:酿酒废酵母对共存重金属离子Pb2+、Ni 2+、Cu2+吸附的最佳条件为pH值3.15,吸附温度30℃,吸附时间60min,酵母浓度2.04g/L,离子浓度61.34mg/L,该条件下酿酒废酵母对共存离子的吸附率为62.78%。     

Simultaneous accumulation of proline and trehalose in industrial baker's yeast enhances fermentation ability in frozen dough

Freeze tolerance is a necessary characteristic for industrial baker's yeast because frozen-dough baking is one of the key technologies for supplying oven-fresh bakery products to consumers. Both proline and trehalose are known to function as cryoprotectants in yeast cells. In order to enhance the freeze tolerance of yeast cells, we constructed a self-cloning diploid baker's yeast strain with simultaneous accumulation of proline, by expressing the PRO1-I150T allele, encoding the proline-feedback inhibition-less sensitive γ-glutamyl kinase, and trehalose, by disrupting the NTH1 gene, encoding neutral trehalase. The resultant strain retained higher tolerance to oxidative and freezing stresses than did the single proline- or trehalose-accumulating strain. Interestingly, our results suggest that proline and trehalose protect yeast cells from short-term and long-term freezing, respectively. Simultaneous accumulation of proline and trehalose in industrial baker's yeast also enhanced the fermentation ability in the frozen dough compared with the single accumulation of proline or trehalose. These results indicate that baker's yeast that accumulates both proline and trehalose is applicable for frozen-dough baking.     

Cell surface hydrophobicity and colony morphology of Trichosporon asahii clinical isolates

Trichosporon asahii is a pathogenic basidiomycetous yeast. Individual strains of T. asahii have different colony morphologies. However, it is not clear whether cell surface phenotypes differ among the colony morphologies. Here we characterized the cell surface hydrophobicity and analysed the carbohydrate contents of the cell surface polysaccharides in T. asahii clinical isolates with various colony morphologies. Among the three distinctive colony morphologies obtained from one clinical isolate, the white‐type morphology exhibited higher hydrophobicity. The hydrophobicity of heat‐killed T. asahii cells was greatly reduced after periodate oxidation of the cell surface carbohydrates. Furthermore, the cell wall and extracellular polysaccharide components differed among the morphologies. Our results suggest that T. asahii cell surface hydrophobicity is affected by cell surface carbohydrate composition. Copyright © 2016 John Wiley & Sons, Ltd.     

霞多丽葡萄自然发酵过程中酵母种群的分布与变化规律

采用WL形态学归类分析和5.8S-ITS rDNA PCR-RFLP分子生物学鉴定方法,对我国2个主要产区所产白葡萄品种霞多丽自然发酵过程中的酵母种群进行了系统研究。结果表明,从霞多丽葡萄发酵液中分离到7个酵母菌种,不同产区霞多丽发酵液中分布酵母菌的种类和数量有一定差别,但在发酵过程中酵母种群的变化趋势是一致的。     

低产乙醛啤酒酵母的定向驯化筛选

为降低啤酒中乙醛含量,采用紫外线对1株啤酒工业生产菌株MI4进行诱变,经双硫仑平板初筛、乙醛培养基驯化复筛,获得了1株低产乙醛的啤酒酵母D-A-14。与出发菌株MI4相比,采用该突变株酿制的啤酒中乙醛含量为2.86 mg/L,降低了76%;且高级醇总量降低而酯含量升高,风味更加协调。这表明筛选得到的低乙醛突变株适于啤酒工业生产。     

Vitamins in brewing: presence and influence of thiamine and riboflavin on wort fermentation

Thiamine and riboflavin vitamers are present in a wide range of foods including beer. These vitamers play critical roles in a variety of enzymatic complexes and can promote and maintain metabolism. Currently, the presence and role of these vitamers in the malting and brewing industry have not been widely explored. This research investigated the effects of various fermentation conditions that may lead to the variations in the vitamin content in beer observed by previous researchers. The present research found that during fermentation, the thiamine content of wort is quickly utilized within the first 6 h of a standard fermentation and the uptake of this vitamin is not affected by increases in wort gravity. While no significant changes were observed in extracellular phosphorylated vitamers of thiamine, both free thiamine and thiamine diphosphate accumulated intracellularly during the wort fermentation. Meanwhile extracellular riboflavin vitamers were only poorly utilized during beer fermentations, however flavin mononucleotide rapidly accumulated intracellularly and more so under aerobic conditions. When yeast was exposed to an all‐malt high‐gravity wort, the thiamine or riboflavin utilization was not affected. However, thiamine utilization was reduced in adjunct‐driven high‐gravity worts. Notwithstanding the lowered thiamine uptake under high‐gravity conditions; there were some minor improvements in fermentation performance and yeast viability. The addition of thiamine to an all‐malt wort did appear to enhance yeast viability, both under normal and high‐gravity conditions. Copyright © 2016 The Institute of Brewing & Distilling     

啤酒酿造酵母原生质体融合株GR5的中试

报道了啤酒酵母原生质体融合株GR5的中试结果。融合株GR5的凝絮性较强 (本斯值为 2 .7) ,以12°Bx麦芽汁为培养基 ,用 5 0 0L发酵罐在 12℃下发酵 ,发酵至第 8d菌株GR5的发酵度为 6 9.2 % ,发酵液中的双乙酰含量为 0 .0 4 98mg/L、乙醛含量为 6 .34mg/L ,总高级醇含量为 74 .4mg/L ,该菌株生产的啤酒 ,口感柔和、协调 ,略带酯香味。融合株GR5的遗传性稳定 ,是一株具有工业应用前景的啤酒酿造酵母菌株