Stringini Marzia, Comitini Francesca, Taccari Manuela, Ciani Maurizio
Dipartimento SAIFET, sez. di Microbiologia Alimentare, Industriale e Ambientale, Università Politecnica delle Marche, via Brecce Bianche, 60131 Ancona, Italy.
Food Microbiol. 2009 Jun;26(4):415-20. doi: 10.1016/j.fm.2009.02.006. Epub 2009 Feb 28.
In the present study, we have investigated the occurrence of yeast flora during tapping and fermentation of palm wine from Cameroon. The yeast diversity was investigated using both traditional culture-dependent and culture-independent methods. Moreover, to characterize the isolates of the predominant yeast species (Saccharomyces cerevisiae) at the strain level, primers specific for delta sequences and minisatellites of genes encoding the cell wall were used. The results confirm the broad quantitative presence of yeast, lactic acid bacteria and acetic acid bacteria during the palm wine tapping process, and highlight a reduced diversity of yeast species using both dependent and independent methods. Together with the predominant species S. cerevisiae, during the tapping of the palm wine the other species found were Saccharomycodes ludwigii and Zygosaccharomyces bailii. In addition, denaturing gradient gel electrophoresis (DGGE) analysis detected Hanseniaspora uvarum, Candida parapsilopsis, Candida fermentati and Pichia fermentans. In contrast to the progressive simplification of yeast diversity at the species level, the molecular characterization of the S. cerevisiae isolates at the strain level showed a wide intraspecies biodiversity during the different steps of the tapping process. Indeed, 15 different biotypes were detected using a combination of three primer pairs, which were well distributed in all of the samples collected during the tapping process, indicating that a multistarter fermentation takes place in this particular natural, semi-continuous fermentation process.
在本研究中,我们调查了喀麦隆棕榈酒采集和发酵过程中酵母菌群的出现情况。使用传统的依赖培养法和不依赖培养法对酵母多样性进行了研究。此外,为了在菌株水平上对主要酵母菌种(酿酒酵母)的分离株进行表征,使用了针对编码细胞壁基因的δ序列和微卫星的特异性引物。结果证实了在棕榈酒采集过程中酵母、乳酸菌和醋酸菌在数量上广泛存在,并突出显示了使用依赖培养法和不依赖培养法时酵母菌种多样性的降低。与主要菌种酿酒酵母一起,在棕榈酒采集过程中发现的其他菌种有路德类酵母和拜耳接合酵母。此外,变性梯度凝胶电泳(DGGE)分析检测到了葡萄汁有孢汉逊酵母、近平滑念珠菌、发酵念珠菌和发酵毕赤酵母。与酵母菌种水平上多样性的逐渐简化相反,酿酒酵母分离株在菌株水平上的分子表征显示在采集过程的不同阶段存在广泛的种内生物多样性。事实上,使用三对引物组合检测到了15种不同的生物型,它们在采集过程中收集的所有样本中分布良好,这表明在这个特定的自然半连续发酵过程中发生了多起始发酵。