Facultad de Ciencias Biológicas, Universidad de Ciencias y Artes de Chiapas, Libramiento Norte 1150, colonia Lajas Maciel, Tuxtla Gutiérrez, México.
Antonie Van Leeuwenhoek. 2011 Nov;100(4):497-506. doi: 10.1007/s10482-011-9605-y. Epub 2011 Jun 17.
The aims of this work were to characterize the fermentation process of mezcal from San Luis Potosi, México and identify the yeasts present in the fermentation using molecular culture-dependent methods (RFLP of the 5.8S-ITS and sequencing of the D1/D2 domain) and also by using a culture-independent method (DGGE). The alcoholic fermentations of two separate musts obtained from Agave salmiana were analyzed. Sugar, ethanol and major volatile compounds concentrations were higher in the first fermentation, which shows the importance of having a quality standard for raw materials, particularly in the concentration of fructans, in order to produce fermented Agave salmiana must with similar characteristics. One hundred ninety-two (192) different yeast colonies were identified, from those present on WL agar plates, by RFLP analysis of the ITS1-5.8S- ITS2 from the rRNA gene, with restriction endonucleases, HhaI, HaeIII and HinfI. The identified yeasts were: Saccharomyces cerevisiae, Kluyveromyces marxianus, Pichia kluyveri, Zygosaccharomyces bailii, Clavispora lusitaniae, Torulaspora delbrueckii, Candida ethanolica and Saccharomyces exiguus. These identifications were confirmed by sequencing the D1-D2 region of the 26S rRNA gene. With the PCR-DGGE method, bands corresponding to S. cerevisiae, K. marxianus and T. delbrueckii were clearly detected, confirming the results obtained with classic techniques.
本工作的目的是对来自墨西哥圣路易斯波托西的梅斯卡尔酒的发酵过程进行表征，并使用分子培养依赖方法（5.8S-ITS 的 RFLP 和 D1/D2 结构域的测序）和非培养方法（DGGE）鉴定发酵过程中存在的酵母。分析了从龙舌兰 salmiana 获得的两个单独醪液的酒精发酵。第一个发酵中的糖、乙醇和主要挥发性化合物浓度较高，这表明对原材料（特别是果聚糖浓度）进行质量标准控制非常重要，以便生产具有类似特性的发酵龙舌兰 salmiana 醪液。通过 rRNA 基因 ITS1-5.8S-ITS2 的 HhaI、HaeIII 和 HinfI 限制性内切酶分析，从 WL 琼脂平板上存在的酵母，鉴定了 192 个不同的酵母菌落。鉴定出的酵母有：酿酒酵母、马克斯克鲁维酵母、毕赤酵母、近平滑假丝酵母、乳酸克鲁维酵母、德巴利酵母、燃料乙醇假丝酵母和矮小假丝酵母。通过对 26S rRNA 基因的 D1-D2 区进行测序，确认了这些鉴定结果。通过 PCR-DGGE 方法，清楚地检测到与 S. cerevisiae、K. marxianus 和 T. delbrueckii 相对应的条带，证实了经典技术获得的结果。
