福建地区葡萄自然发酵过程中酵母菌的组成及差异分析

Composition and difference of yeast populations during spontaneous fermentation of different varieties of grapes from Fujian

  • 摘要: 【目的】对福建地区葡萄自然发酵酵母菌组成的差异进行分析,明确福建主要酵母菌种类,为酿制具有南方地域特色的葡萄酒提供优良菌株。【方法】采集夏黑、刺葡萄和玫瑰香葡萄自然发酵不同阶段发酵前期(7 d)、中期(15 d)和后期(25 d)的发酵液进行酵母菌分离,采用WL培养基进行酵母菌表型性状鉴别,挑取代表性菌株进行DNA提取及26S rDNA D1/D2鉴定,分析不同发酵阶段酵母菌的组成差异。通过测定相同发酵条件下的酒精度、总糖、还原糖和总酸含量,对酿酒酵母的酿酒性能进行初步分析。【结果】不同品种葡萄自然发酵的发酵液中共分离获得114株菌株,根据其表型鉴定为18个不同的表型类群。进一步通过酵母菌的26S r DNA D1/D2区序列测序,可将114株酵母菌菌株划分为18种,包括拜耳接合酵母(Zygosaccharomyces bailii)、双孢接合酵母(Zygosaccharomyces bisporus)、酿酒酵母(Saccharomyces cerevisiae)等。不同品种葡萄发酵过程中酵母菌种类和组成具有一定的差异,其中刺葡萄A有10种、刺葡萄B有11种、夏黑A和玫瑰香均为7种、夏黑B有8种。陆生伊萨酵母(Issatchenkia terricola)出现在刺葡萄A、玫瑰香和夏黑B发酵前期,且所占比例最高,分别达100.00%、50.00%和40.00%;盔形毕赤酵母(Pichia galeiformis)是夏黑A和刺葡萄B发酵前期的优势菌;在不同品种葡萄自然发酵中期均含有泽普林假丝酵母(Candida zemplinina)和盔形毕赤酵母;刺葡萄A、刺葡萄B和夏黑B发酵后期的优势菌为盔形毕赤酵母,所占比例分别为40.00%、55.56%和71.43%;酿酒酵母在夏黑A、玫瑰香和夏黑B发酵中期开始出现,直到发酵后期,均占有较大比例,分别为66.67%、42.86%和14.29%。初步葡萄酿酒发酵表明,酿酒酵母VMH-M042的酿酒性能(酒精度12.5%、总糖含量76.0 g/L、还原糖含量40.5 g/L、总酸含量5.6%)优良。【结论】福建地区不同品种葡萄分离得到的酵母菌资源丰富,区域性差异明显,特异性资源较多,其中酿酒酵母VMH-M042在葡萄酒发酵中表现出较优良的酿酒性能,可为南方葡萄酒酿制提供丰富的菌种资源和理论依据。

     

    Abstract: 【Objective】In order to provide good yeast strains for wine brewing with the characteristics of southern region, the difference of the yeast populations were analyzed and the main yeast species were identifiedin the spontaneous fermentation of grapes from Fujian.【Method】The yeast strains were isolated from fermentation of Xiahei, Vitis davidiiFoex. and Muscat Hamburg at different stagesearlyfermentation stage(7 d), middle fermentation stage(15 d), later fermentation stage(25 d) . The strains were preliminarily classified based on WL nutrient agar, DNA was extracted and 26 S rDNA D1/D2 sequence analysis was conducted on representative strains. Analysis of the composition differences in yeast across the fermentation stages was conducted. The brewing performance of Saccharomyces cerevisiae was analyzed by determine the alcohol content, total sugar, reducing sugar and total acid under the same fermentation conditions.【Result】A total of 114 strains of yeast were isolated from fermentation liquid of different varieties and preliminarily classified as 18 phenotypic groups. The 26 S r DNA D1/D2 sequence analysis showed that they were identified as 18 different known yeast species, such as Zygosaccharomyces bailii, Z. bisporus and S. cerevisiae, etc. The composition of yeast populations in different stages of natural fermentation of different varieties of grapes was different. There were 10 kinds in V. davidii Foex.A, 11 kinds in V. davidii Foex.B, 7 kinds in Xiahei A, 8 kinds in Xiahei B, and 7 kinds in Muscat Hamburg.In the early stage of the fermentation of V. davidii Foex. A, Muscat Hamburg and Xiahei B, the preponderant microzymes were Issatchenkia terricola with the highest proportions. The proportions were 100.00%, 50.00% and 40.00%, respectively. In the early stage of the fermentation of Xiahei A and V. davidii Foex.B, the preponderant microzymes were Pichia galeiformis. Candida zemplinina and P. galeiformis were involved in the middle stage of the different varieties of grapes. At the late stage of the fermentation of V. davidii Foex. A, Muscat Hamburg and Xiahei B, the preponderant microzyme was P. galeiformis, whose proportions were 40.00%, 55.56% and 71.43%, respectively. S. cerevisiae was involved in both of the middle and the late stages of the fermentation of Xiahei A, Muscat Hamburg and Xiahei B. Its proportions at the late stage were 66.67%, 42.86% and 14.29%. Preliminary studies of grape fermentation showed the enological characteristics(alcohol content 12.5%, total sugar 76.0 g/L, reducing sugar 40.5 g/L, and total acid 5.6%) of the S. cerevisiae strain VMH-M042 were excellent.【Conclusion】The yeast resources isolated from different grape varieties in Fujian region are abundant with obvious regional differences and more specific resources. Among them, S. cerevisiae strain VMH-M042 has good fermentation performance. Studying on the composition and difference of yeast populations in different stages of spontaneous fermentation of different varieties of grapes from southern region can provide more yeast strain resources and lay a foundation for the local wine brewing.

     

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