Margalef-Català Mar, Araque Isabel, Bordons Albert, Reguant Cristina
Grup de Biotecnologia Enològica, Departament de Bioquímica i Biotecnologia, Facultat d'Enologia, Universitat Rovira i Virgili, Marcel·lí Domingo, 1, 43007 Tarragona, Catalonia, Spain.
Grup de Biotecnologia Enològica, Departament de Bioquímica i Biotecnologia, Facultat d'Enologia, Universitat Rovira i Virgili, Marcel·lí Domingo, 1, 43007 Tarragona, Catalonia, Spain.
Int J Food Microbiol. 2017 Feb 2;242:61-69. doi: 10.1016/j.ijfoodmicro.2016.11.013. Epub 2016 Nov 16.
Although Oenococcus oeni is the main species that is responsible for malolactic fermentation (MLF), harsh wine conditions can limit its performance. Although several mechanisms underlying the response to stress have been studied in this species, little is known regarding the cellular systems that protect against oxidative stress in other bacteria, such as glutathione (GSH). O. oeni cannot synthesize GSH but contains several genes related to its utilization. In this study, the relative expression (RE) of the seven genes involved in the GSH redox system found in O. oeni PSU-1 (gshR, gpo, three glutaredoxin-like genes and two subunits of an hypothetical transporter) has been measured. The study was performed using three strains, with each exhibiting a different GSH uptake capacity. The strains were grown in a stress-adaptation medium supplemented with 5mM GSH and under different adaptation stress conditions (pH4 and 6% ethanol). The RE showed that only some of these genes, including one for a possible glutaredoxin (OEOE_RS04215) and cydC for a subunit of a putative GSH transporter (OEOE_RS1995), responded to the addition of GSH. The presence of ethanol had a relevant effect on gene expression. Among the studied genes, the one for a NrdH-redoxin (OEOE_RS00645) showed a common response to ethanol in the strains, being over-expressed when grown with GSH. In most cases, the transcriptional changes were more evident for the strain with a higher capacity of GSH uptake. Malolactic performance of the three strains after pre-adaptation was evaluated in wine-like media (12% ethanol and pH3.4). It was observed that the addition of GSH during pre-adaptation growth had a protective role in the cells exposed to low pH and ethanol, resulting in a quicker MLF.
尽管酒酒球菌是负责苹果酸-乳酸发酵(MLF)的主要菌种,但恶劣的葡萄酒环境会限制其性能。虽然已经对该菌种应对压力的几种潜在机制进行了研究,但对于其他细菌中抵御氧化应激的细胞系统,如谷胱甘肽(GSH),却知之甚少。酒酒球菌无法合成GSH,但含有几个与其利用相关的基因。在本研究中,测定了酒酒球菌PSU-1中发现的参与GSH氧化还原系统的七个基因(gshR、gpo、三个类谷氧还蛋白基因和一个假定转运蛋白的两个亚基)的相对表达(RE)。该研究使用了三株菌株,每株表现出不同的GSH摄取能力。这些菌株在添加了5mM GSH的应激适应培养基中,并在不同的适应应激条件(pH4和6%乙醇)下生长。RE表明,这些基因中只有一些,包括一个可能的谷氧还蛋白基因(OEOE_RS04215)和一个假定GSH转运蛋白亚基的cydC基因(OEOE_RS1995),对添加GSH有反应。乙醇的存在对基因表达有显著影响。在所研究的基因中,一个NrdH-氧化还原蛋白基因(OEOE_RS00645)在菌株中对乙醇表现出共同反应,在与GSH一起生长时过度表达。在大多数情况下,转录变化对于具有较高GSH摄取能力的菌株更为明显。在类似葡萄酒的培养基(12%乙醇和pH3.4)中评估了三株菌株预适应后的苹果酸-乳酸发酵性能。观察到在预适应生长期间添加GSH对暴露于低pH和乙醇的细胞具有保护作用,从而导致更快的MLF。