中国农业科学 ›› 2012, Vol. 45 ›› Issue (19): 4067-4074.doi: 10.3864/j.issn.0578-1752.2012.19.019

• 园艺 • 上一篇    下一篇

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   

  1. 1.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
  • 收稿日期:2012-03-27 出版日期:2012-10-01 发布日期:2012-08-21
  • 通讯作者: 通信作者刘延琳,E-mail:yanlinliu@nwsuaf.edu.cn
  • 作者简介:马 捷,E-mail:chevalblanc1986@gmail.com
  • 基金资助:

    国家葡萄产业技术体系建设专项经费(CARS-30-jg-03)、国家自然科学基金项目(31271917)

Construction of Wine Yeast for Improving Cystathionine β-Lyase Activity

 MA  Jie, LIU  Yan-Lin   

  1. 1.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
  • Received:2012-03-27 Online:2012-10-01 Published:2012-08-21

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&beta, -裂解酶, 硫醇

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 of engineering yeast strains was one-day longer, the residual sugars and acetic acid were higher than their host strain, while the production of thiols increased obviously, which were 2.8-4.3 folds than that in the host strain. In Sauvignon Blanc wine making test, the technological characteristics of engineering yeast strains showed no significant difference with the host strain, whereas the production of thiols was 1.22 times higher than that of the host strains.【Conclusion】The cystathionine β-Lyase activity of wine yeast was enhanced successfully by using the gene modification method in this study. It will have great interests for improving flavor characteristics of wine in the future.

Key words: Saccharomyces cerevisiae, cystathionine β-Lyase, thiols