Molecular and biochemical aspects of Brettanomyces in brewing

Brettanomyces is a semi‐domesticated yeast that is a crucial component of lambic beers and is increasingly attracting the attention of the brewing industry. Brettanomyces display Saccharomyces‐like features, such as a positive Crabtree effect, ethanol synthesis and tolerance to harsh environments. Additionally, Brettanomyces exhibit β‐glucosidase and esterase activities, the production of phenolic compounds and tetrahydropyridines, together with the ability to ferment dextrins and breakdown cellobiose from wooden casks. Although the importance of Brettanomyces species is documented in the production of different beer styles, the molecular and biochemical features of these species required for brewing are poorly understood. Therefore, this work reviews the current knowledge of the molecular biology and biochemistry underlying the performance of Brettanomyces in the brewing industry. © 2019 The Institute of Brewing & Distilling     

Monovalent cation transporters at the plasma membrane in yeasts

Maintenance of proper intracellular concentrations of monovalent cations, mainly sodium and potassium, is a requirement for survival of any cell. In the budding yeast Saccharomyces cerevisiae, monovalent cation homeostasis is determined by the active extrusion of protons through the Pma1 H⁺-ATPase (reviewed in another chapter of this issue), the influx and efflux of these cations through the plasma membrane transporters (reviewed in this chapter), and the sequestration of toxic cations into the vacuoles. Here, we will describe the structure, function, and regulation of the plasma membrane transporters Trk1, Trk2, Tok1, Nha1, and Ena1, which play a key role in maintaining physiological intracellular concentrations of Na⁺, K⁺, and H⁺, both under normal growth conditions and in response to stress.     

Inactivation behaviors of foodborne microorganisms in multi-frequency power ultrasound-treated orange juice

This study established the inactivation kinetic parameters of some pathogenic bacteria including Escherichia coli O157:H7, Salmonella enterica serotypes, and Listeria monocytogenes; and spoilage yeasts namely, Debaryomyces hansenii, Clavispora lusitaniae, Torulaspora delbrueckii, Pichia fermentans, and Saccharomyces cerevisiae in orange juice subjected to multi-frequency Dynashock power ultrasound treatment. All test organisms exhibited a biphasic inactivation behavior with a sigmoidal inactivation curve consisted of an initial inactivation lag, followed by logarithmic linear inactivation. Injury accumulation in the inactivation lag phase was established in acid-adapted bacteria. The time necessary to reduce initial inoculated populations by 5 log cycles (99.999%), T_5D values, significantly increased with acid adaptation. The T_5D of E. coli, S. enterica, and L. monocytogenes increased from 37.64, 36.87, and 34.59 respectively; to 54.72, 40.38, and 37.83 min respectively after acid exposure. Temperature increase due to sensible heat propagation during ultrasound treatment decreased the resistance of the test bacteria. The cocktail of E. coli O157:H7 had significantly greater resistance towards ultrasound treatment (T_5D = 54.72 min) than any of the individual strain (T_5D = 41.48–47.48 min) in the mix. Similar results were found in the composited (T_5D = 60.02 min) and individual species (T_5D = 20.31–59.04 min). The results established in this work provide baseline information on microbial behavior in multi-frequency ultrasound-treated orange juice for establishment of pasteurization process schedules.     

The Production of Low-Alcohol Wines by Aerobic Yeasts

Aerobic yeasts of the genera Pichia and Williopis are commonly regarded as spoilage yeasts of beer and wine by causing turbidity, a surface film of yeast growth and often an excessive estery flavour. However, their ability to utilise sugars oxidatively for cell growth with the production of estery and other flavours of wine with only minimal production of ethanol suggests a method for the production of low-alcohol wines of pleasant “fermented” flavour without the need for additional equipment to remove alcohol by dialysis, reverse osmosis or distillation, or without the excessive sweetness remaining from arrested fermentation. Three strains of Pichia and one of Williopsis were examined for their ability to produce approximately 3%(v/v) ethanol and a good estery and fruity flavour. With normal anaerobic fermentation conditions, or with gentle stirring to prevent formation of a surface film, excessive amounts of alcohol were produced from grape juice of 15% or 20% (w/v) initial sugar concentration. However, an acceptably flavoured wine of alcohol content < 3% was produced by agitation and aeration during fermentation. The ethanol formed in the early stages of culture was oxidised to a final level < 3%, with the production of cell mass and an acceptable flavour.     

橙汁腐败酵母菌的分子鉴定及其形态特征分析

目的：结合多种酵母菌鉴定方法,分析腐败橙汁中酵母菌的种类,为快速检测和控制商品橙汁中酵母菌污染奠定基础,也为酵母菌种类的快速鉴定提供参考。方法：以分离自腐败橙汁的8株酵母菌株为材料,观察菌落和细胞形态。用引物ITS1/ITS4扩增菌株5.8S rDNA 及其ITS间隔区(5.8S-ITS),对扩增产物用HhaⅠ、Hae Ⅲ 和 HinfⅠ进行限制性酶切片段多态性(RFLP)分析和测序;用引物NL1/NL4扩增26S rDNA D1/D2区并测序。结果：菌落特征观察将菌株初分为6组,细胞学观察粗分为3组,分子方法都分成4组。用限制性内切酶HhaⅠ、Hae Ⅲ和HinfⅠ对5.8S-ITS区产物进行 RFLP分析,观察到4种不同的图谱类型。5.8S-ITS 区和26S rDNA D1/D2区的序列分析结果相似,8株分离菌与GenBank中的4种酵母菌参考菌株序列一致性达99%以上。分离株与参考菌株以两区序列构建的NJ系统树都分成4枝：Y1与克鲁维毕赤酵母(Pichia kluyveri),Y12-3、Y18-1与发酵毕赤酵母(Pichia fermentans),Y22、Y23、Y26和Y56与Meyerozyma guilliermondii,Y47与Wickerhamomyces anomalus分别聚为一枝。结论：结合形态分析和核酸分析,将8株腐败橙汁分离菌鉴定为P. kluyveri var. kluyveri、P. fermentans、M. guilliermondii和W. anomalus 4个种,其中M. guilliermondii在腐败橙汁中首见报道;ITS- RFLP分析、26S rDNA D1/D2区测序与菌株形态特征结合能有效鉴定酵母菌,核糖体DNA分析可鉴定酵母到生物学种,菌落形态特征可反映种以下的遗传差异,因此,采用分子方法鉴定时不能忽视形态特征分析的重要性。