Univ. Bourgogne Franche-Comté, AgroSup Dijon, PAM UMR A 02.102, F-21000, Dijon, France.
Univ. Bourgogne Franche-Comté, AgroSup Dijon, PAM UMR A 02.102, F-21000, Dijon, France.
Food Microbiol. 2016 Dec;60:21-8. doi: 10.1016/j.fm.2016.06.012. Epub 2016 Jun 16.
Besides deacidifying wine, Oenococcus oeni bring significant changes in the chemical composition of wine by releasing esters by the action of their own esterases. The impact of O. oeni esterases remains relatively unexplored. Four esterase genes were identified from O. oeni genome (estA2, estA7, estC, and estB). The dual objective of this study was, first to use a genetic tool enabling the expression of esterase genes in enological conditions and, second, to investigate the impact of O. oeni esterase gene expression during winemaking on wine aromatic profile. Both estA2 and estA7 genes were successfully cloned and expressed in O. oeni and recombinant strains were inoculated in Aligoté wine to initiate malolactic fermentation (MLF). Ester profile of experimental wine was established by SPME-GC-MS. EstA2 caused significant decreases in the concentrations of isoamyl acetate, ethyl hexanoate, isobutyl acetate, and hexyl acetate, by 42.7%, 23.4%, 51.5%, and 28.9%, respectively. EstA2 has preferential hydrolytic activity toward acetate esters from higher alcohols. EstA7 has synthetic activity toward hexyl acetate with a significant 22.7% increase. This study reports the first efficient expression system enabling the production of a functional protein in O. oeni in enological conditions.
除了降低葡萄酒的酸度外,酒酒球菌通过自身酯酶的作用释放酯类物质,从而使葡萄酒的化学成分发生显著变化。酒酒球菌酯酶的影响仍相对未知。从酒酒球菌基因组中鉴定出了 4 个酯酶基因(estA2、estA7、estC 和 estB)。本研究的双重目的是,首先使用一种遗传工具,使酯酶基因在酿造条件下表达,其次研究在葡萄酒酿造过程中表达酒酒球菌酯酶基因对葡萄酒芳香特征的影响。成功地克隆了 estA2 和 estA7 基因并在酒酒球菌中表达,重组菌株被接种到阿里高特葡萄酒中启动苹果酸-乳酸发酵(MLF)。通过 SPME-GC-MS 建立了实验葡萄酒的酯类图谱。EstA2 导致异戊酸乙酯、己酸乙酯、乙酸异丁酯和乙酸己酯的浓度分别显著降低了 42.7%、23.4%、51.5%和 28.9%。EstA2 对来自高级醇的乙酸酯具有优先的水解活性。EstA7 对己酸乙酯具有合成活性,增加了 22.7%。本研究报道了第一个有效的表达系统,能够在酿造条件下在酒酒球菌中生产功能性蛋白质。