Coucheney Françoise, Desroche Nicolas, Bou Magali, Tourdot-Maréchal Raphaëlle, Dulau Laurent, Guzzo Jean
Laboratoire de Microbiologie, UMR Université de Bourgogne/INRA 1232, ENSBANA, 1, Esplanade Erasme, 21 000 Dijon, France.
Int J Food Microbiol. 2005 Dec 15;105(3):463-70. doi: 10.1016/j.ijfoodmicro.2005.04.023. Epub 2005 Aug 2.
The lactic acid bacterium Oenococcus oeni, mainly responsible for malolactic fermentation (MLF), is used in new winery process as starter culture for direct inoculation. The difficulty to master MLF according to the wine led us to search a new approach to select effective O. oeni strains. Biochemical and molecular tests were performed in order to characterize three strains of O. oeni selected for malolactic starter elaboration. Malolactic and ATPase activities that appeared as a great interest in MLF were measured and the expression of a small heat shock protein Lo18 was evaluated by immunoblotting and real-time PCR. These results were correlated with the performances of strains in two red wines. Physiological and molecular characteristics of the three strains showed significant differences for the global malolactic activity on intact cell at pH 3.0 and at the level of induction of the small heat shock protein Lo18. These two parameters appeared of interest to evaluate in the ability of O. oeni strains to survive into wine after direct inoculation and to perform MLF. Indeed, a tested strain that presented the highest malolactic activity on intact cells at pH 3.0 and a high level of Lo18 induction showed a high growth rate and a high specific kinetic of malate consumption. The techniques used in this work carry out more quickly and more reliable than usual for the selection of effective strains intended for direct inoculation in wines.
乳酸乳球菌,主要负责苹果酸 - 乳酸发酵(MLF),在新的酿酒工艺中用作直接接种的起始培养物。根据葡萄酒来掌控MLF存在困难,这促使我们寻找一种新方法来筛选有效的乳酸乳球菌菌株。为了对三种用于制备苹果酸 - 乳酸发酵起始培养物而挑选出的乳酸乳球菌菌株进行表征,我们进行了生化和分子测试。测定了在MLF中表现出极大研究价值的苹果酸 - 乳酸酶活性和ATP酶活性,并通过免疫印迹和实时PCR评估了小热休克蛋白Lo18的表达。这些结果与菌株在两种红葡萄酒中的性能相关。这三种菌株的生理和分子特征在pH 3.0时完整细胞的总体苹果酸 - 乳酸活性以及小热休克蛋白Lo18的诱导水平上显示出显著差异。这两个参数对于评估乳酸乳球菌菌株在直接接种到葡萄酒后在其中存活并进行MLF的能力似乎具有重要意义。事实上,在pH 3.0时对完整细胞表现出最高苹果酸 - 乳酸活性且Lo18诱导水平较高的受试菌株显示出高生长速率和高苹果酸消耗比动力学。这项工作中使用的技术比通常用于筛选用于直接接种到葡萄酒中的有效菌株的方法执行得更快且更可靠。