具有高胱硫醚beta;-裂解酶活性的葡萄酒酵母工程菌株的构建.pdfVIP

中国农业科学 2012,45(19):4067-4074 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2012.19.019 具有高胱硫醚 β-裂解酶活性的葡萄酒酵母工程菌株的构建 马 捷，刘延琳 （西北农林科技大学葡萄酒学院/ 陕西省葡萄与葡萄酒工程技术研究中心，陕西杨凌 712100 ） 摘要：【目的】构建具有较高胱硫醚β-裂解酶活性的葡萄酒酵母工程菌株，用于提高葡萄酒的香气品质。【方 法】利用质粒 pAUR123 将大肠杆菌 tnaA 基因在中国本土酿酒酵母菌株 LFP525 中进行表达，得到的转化子进行酶 活测定，并通过模拟汁和干白的发酵，评价其酿酒特性和产生硫醇类物质的能力。【结果】成功获得酵母工程菌株 TH1和TH2，其胱硫醚β-裂解酶活性与受体菌比提高了32.25%-59.44%。模拟葡萄汁发酵显示,工程菌株中硫醇类 物质的产量显著提高，是受体菌的 2.8-4.3 倍。工程菌株的发酵时间比受体菌略长，残糖和挥发酸显著高于宿主 菌，但其含量处于正常范围。在长相思干白发酵试验中，该工程菌株的主要酿酒特性与受体菌相差不大，且硫醇 类物质的产量提高了1.22倍。【结论】通过基因改造可以提高酿酒酵母胱硫醚β-裂解酶活性，应用改良菌株可进 一步提高葡萄酒的香气品质。 关键词：酿酒酵母；胱硫醚β-裂解酶；硫醇 Construction of Wine Yeast for Improving Cystathionine β-Lyase Activity MA Jie, LIU Yan-lin (College of Enology，Northwest A ＆F University/Shaanxi Engineering Research Center for Viti-viniculture, Yangling 712100, Shaanxi) Abstract: 【Objective 】 The wine yeast which exhibited higher cystathionine β-Lyase activity was contructed. 【Method 】 Plasmid pAUR123 was used to express the tnaA gene of Escherichia coli in Saccharomyces cerevisiae strain LFP525 which is a indigenous wine yeast selected from China. Then the cystathionine β-Lyase activity of yeast transformants was detected. After that, model grape juice fermentation and white wine fermentation were carried out to evaluate the tenological properties and thiol-producing characteristics of them. 【Result 】Two engineering wine yeasts, TH1 and TH2, were obtained in this study. Compared with the host strain, their cystathionine β-Lyase activities were increased by 32.25% to 59.44%. The result of model grape juice fermentation showed that the fermentation duration