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首页> 外文期刊>Journal of the Institute of Brewing >Use of PCR-DHPLC (Polymerase Chain Reaction-Denaturing High Performance Liquid Chromatography) for the Rapid Differentiation of Industrial Saccharomyces pastorianus and Saccharomyces cerevisiae Strains
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Use of PCR-DHPLC (Polymerase Chain Reaction-Denaturing High Performance Liquid Chromatography) for the Rapid Differentiation of Industrial Saccharomyces pastorianus and Saccharomyces cerevisiae Strains

机译：PCR-DHPLC（聚合酶链反应-变性高效液相色谱法）用于快速分离工业酿酒酵母和酿酒酵母菌株的方法

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摘要

Strain specific detection and control of Saccharomyces pastorianus and Saccharomyces cerevisiae starter cultures is of great importance for the fermentation industry. The preconditions of strain specific fermentation characteristics can be ensured by periodic analysis and confirmation of the strain identity. With regard to industrial S. pastorianus and S. cerevisiae strains and a focus on brewing strains, the differentiation methods most available are time-consuming and not very discriminative. In this work PCR-DHPLC analysis was investigated as a novel approach for the differentiation of industrially used S. pastorianus and S. cerevisiae strains. The PCR-DHPLC-system was specific for S. cerevisiae strains and S. pastorianus hybrid strains that contain IGS2 rDNA, which originates from the S. cerevisiae ancestor. For the DNA of 177 strains of 41 non-target species, which are typical for beverage and fermentation surroundings, the absence of PCR-amplificates could be confirmed by DHPLC analysis. It was shown that single strains of S. cerevisiae and S. pastorianus could be differentiated. A strain specific differentiation within the group of top-fermenting Saccharomyces cerevisiae strains could also be performed. For the group of bottom fermenting S. pastorianus brewing strains, strain-to-strain specific differences in the DHPLC chromatograms could be observed which can be used to differentiate and to compare two single strains with each other, although the comparison of chromatograms of an unknown S. pastorianus strain with a set of known S. pastorianus chromatograms could only reveal tendencies towards grouping into types. The differential DHPLC chro-matogram characteristics (fluorescence intensities, number of peaks/side-peaks/peak-shoulders) within S. pastorianus are present, but not as distinctive as for S. cerevisiae. Additionally PCR-DHPLC has advantages compared to other differentiation methods, such as species specificity, speed (2.5 h for one sample) and precision with the described limits.
机译：巴斯德酵母和酿酒酵母发酵剂的菌株特异性检测和控制对于发酵工业具有重要意义。可以通过定期分析和确认菌株身份来确保菌株特异性发酵特性的前提。关于工业巴氏酵母和酿酒酵母菌株以及以酿造菌株为重点，最可用的分化方法是耗时的并且不是非常有区别的。在这项工作中，对PCR-DHPLC分析进行了研究，作为区分工业使用的巴氏酵母和酿酒酵母菌株的一种新方法。 PCR-DHPLC系统对酿酒酵母菌株和含有IGS2 rDNA的酿酒酵母杂交菌株具有特异性，该IGS2 rDNA来源于酿酒酵母祖先。对于饮料和发酵环境中典型的41种非靶标物种的177个菌株的DNA，可以通过DHPLC分析确认不存在PCR扩增子。结果表明，可以区分酿酒酵母和巴斯德酵母的单一菌株。还可以在顶级发酵酿酒酵母菌株组内进行菌株特异性分化。对于一组底部发酵的巴氏毕赤酵母酿造菌株，可以观察到DHPLC色谱图中的菌株间差异，这些差异可用于区分和比较两个单一菌株，尽管未知色谱图的比较带有一组已知的巴氏巴氏酵母色谱图的巴氏巴氏酵母菌株只能显示出归类为类型的趋势。存在巴氏链球菌内的差异DHPLC色谱图特征（荧光强度，峰数/峰/侧峰/峰肩数），但不像酿酒酵母那样独特。此外，与其他区分方法相比，PCR-DHPLC具有优势，例如物种特异性，速度（一个样品为2.5 h）和在上述限制范围内的精密度。

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  • 来源
    《Journal of the Institute of Brewing》 |2010年第4期|p.464-474|共11页
  • 作者

    M. Hutzler; E. Geiger; F. Jacob;

  • 作者单位

    Technische Universitaet Muenchen, Research Centre Weihenstephan for Brewing and Food Quality, Alte Akademie 3, 85354 Freising;

    Technische Universitaet Muenchen, Wissenschaftszentrum Weihen- stephan, Lehrstuhl fuer Technologie der Brauerei II, Alte Akademie 3, 85354 Freising;

    Technische Universitaet Muenchen, Research Centre Weihenstephan for Brewing and Food Quality, Alte Akademie 3, 85354 Freising;

  • 收录信息 美国《科学引文索引》(SCI);
  • 原文格式 PDF
  • 正文语种 eng
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