【目的】高级醇是白酒微量香气成分中的重要组成部分,但是高级醇含量过高会对酒的品质产生不利影响,而白酒中的高级醇主要是在酒精发酵过程由酵母产生的,因此酿酒酵母高级醇合成相关蛋白的研究对于控制高级醇产生具有重要意义。【方法】以诱变得到的低产高级醇酿酒酵母菌株ARTP5和原始酿酒酵母菌株CF4为研究对象,比较两株酵母的胞内蛋白组差异,寻找高级醇合成相关蛋白。【结果】与原始菌株CF4相比,诱变菌株ARTP5高级醇产量降低了20%,有45个胞内蛋白表达量差异2倍以上,通过MALDITOF-MS质谱鉴定出29个,主要包括碳源和能量代谢、胁迫反应过程、蛋白翻译和折叠过程、氨基酸代谢和高级醇代谢等途径的蛋白,其中ARTP5菌株表达上调的支链氨基酸代谢合成途径的ILV5蛋白和表达下调的高级醇合成途径中的ADH1蛋白与诱变菌株ARTP5高级醇降低具有一定相关性。【结论】与高级醇合成具有一定相关性蛋白的发现对于酿酒酵母酒精发酵过程高级醇的合成机制的解析以及白酒酿造过程高级醇产量的控制有重要意义。
[Objective] Higher alcohols is an important part of liquor trace aroma components, but higher alcohols will have negative effects on wine flavor as well as the quality when its content is too high. Higher alcohols in liquor are mainly produced by yeast during alcoholic fermentation and alcoholic fermentation is mainly processed by Saccharomyces cerevisiae, it means that looking for higher alcohol synthesis functional proteins is of great importance for liquor production. [Methods] Two-dimensional electrophoresis was used to compare the differences of intracellular proteins between the Saccharomyces cerevisiae mutant ARTP5, whose higher alcohols production is much less than the origin strain, and the origin strain CF4. Comparative analysis of the two strains' intracellular proteomic differences helps to find higher alcohols synthesis functional proteins. [Results] Compared with the origin strain CF4, the higher alcohols production of mutant strain ARTP5 was reduced by 20% and it indicates that 45 spots (expression ratio≥2) were expressed differently. Among them, 29 proteins were identified by MALDITOF MS-mass spectrometry, including proteins in carbohydrate and energy metabolism, stress response, protein translation and folding, amino acids metabolism, higher alcohol metabolism progress, etc. It is amazing to find out that the protein ILV5 involved in branched chain amino acid metabolism was up-regulated and the protein ADH1 involved in higher alcohol synthesis was down-regulated in strain ARTP5 compared with those in strain CF4, this could possibly be the reason that the higher alcohols production decreased in strain ARTP5. [Conclusion] The finding of the higher alcohols synthesis related protein ILV5 and ADH1 has important significance for the revealing of higher alcohol synthesis mechanism in Saccharomyces cerevisiae alcohol fermentation process.