Synergetic effect of combining PEF treatments with sublethal doses of SO2 on the inactivation of Saccharomyces bayanus and Brettanomyces bruxellensis in red wine

C. Delso, A. Berzosa, Jorge Sanz, Ignacio Álvarez, J. Raso
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Abstract

Certain microorganisms are capable of proliferating in wine despite its low pH and high ethanol content. The yeasts of the Saccharomyces genus responsible for alcoholic fermentation can alter wines with residual sugars; the proliferation of Brettanomyces bruxellensis brings about thoroughly unpleasant sensory changes. The main strategy currently applied in wineries for microbial control is the addition of sulfites (SO2). However, sulfites are being researched due to the symptoms they can cause in allergic individuals. Pulsed electric field (PEF) technology has the capability of inactivating vegetative cells of microorganisms at non-lethal temperatures and could thus prove to be an alternative to SO2. In this study, the resistance of Saccharomyces bayanus and B. bruxellensis suspended in wine to a series of different PEF treatments (10–25 kV/cm; 25–1000 µs; 40–170 kJ/kg) combined with sublethal concentrations of SO2 (10, 25, and 50 ppm) was evaluated. The results showed that even the least intense PEF treatments (10 kV/cm; 115 kJ/kg) inactivated more than 4.0 Log10 cycles in both types of yeasts immediately after treatment. The subsequent incubation of the treated yeasts for 24 h in wine managed to increase inactivation by 3.0 Log10 cycles. The combination of a moderate PEF treatment with sublethal doses of SO2 had a synergistic lethal effect on the two yeasts under study after 24 h of incubation in wine, leading to counts lying below the detection limit (>5.0 Log10 cycles). This synergistic effect was attributed to the existence of a portion of the population that had been sublethally damaged by PEF and in which SO2 could more easily penetrate the cytoplasm. These results demonstrate the capacity of PEF technology for microbial control of spoilage yeasts in wine. PEF could thus represent an alternative with the potential of eliminating or reducing SO2 levels in the winemaking process.
PEF处理与亚致死剂量SO2联合对红葡萄酒中巴亚努斯酿酒酵母和布鲁谢酿酒酵母灭活的协同作用
尽管葡萄酒的pH值低,乙醇含量高,但某些微生物仍能在葡萄酒中繁殖。负责酒精发酵的酵母菌属酵母可以改变含残糖的葡萄酒;bruxellensis Brettanomyces的增殖带来了完全不愉快的感觉变化。目前在酿酒厂中应用的主要微生物控制策略是添加亚硫酸盐(SO2)。然而,亚硫酸盐正在被研究,因为它们会引起过敏个体的症状。脉冲电场(PEF)技术具有在非致死温度下使微生物的营养细胞失活的能力,因此可以证明是二氧化硫的替代品。在本研究中,研究了酒中悬浮的bayanus Saccharomyces和bruxellb对一系列不同PEF处理(10-25 kV/cm;25 - 1000µ年代;40-170千焦/千克)以及二氧化硫亚致死浓度(10、25和50 ppm)进行了评估。结果表明,即使是最弱的PEF处理(10 kV/cm;115 kJ/kg)在处理后立即在两种类型的酵母中灭活超过4.0 Log10循环。随后将处理过的酵母在葡萄酒中孵育24小时,使失活率提高了3.0 Log10个周期。中等PEF处理与亚致死剂量SO2的组合在葡萄酒中孵育24小时后,对所研究的两种酵母具有协同致死作用,导致计数低于检测限(>5.0 Log10周期)。这种协同效应归因于部分种群受到PEF的亚致命性损伤,其中SO2更容易穿透细胞质。这些结果证明了PEF技术对葡萄酒中腐败酵母菌的微生物控制能力。因此,PEF可以代表一种替代方案,具有消除或减少葡萄酒酿造过程中二氧化硫水平的潜力。
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