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Development of an S-layer gene-based PCR-DGGE assay for monitoring dominant Lactobacillus helveticus strains in natural whey starters of Grana Padano cheese

机译:基于S层基因的PCR-DGGE分析方法的开发,用于监测Grana Padano奶酪的天然乳清发酵剂中的瑞士乳杆菌(Lactobacillus helveticus)优势菌株。

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摘要

Monitoring L. helveticus strain dynamics in natural whey starters is of great interest at the industrial level due to the key role that this bacterial population plays in Grana Padano cheese production. In this study, we aimed to develop a PCR-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) assay based on the slpH locus, in parallel with performing culture-dependent analysis of whey samples using optimized media to maximize the number of isolated strains. We designed new primers targeting the slpH locus to amplify a gene region that would be suitable for PCR-DGGE analysis and discriminating strains. Our results confirmed that the developed PCR-DGGE method was rapid and reliable for monitoring the L. helveticus population in whey starter cultures. All sequences of bands detected in the PCR-DGGE profiles from whey samples showed high similarity to S-layer genes of I. helveticus, and perfectly matched with the slpH locus sequences of dominant strains. Overall, our findings indicated that the target region of the slpH locus was sufficiently heterologous to discriminate L. helveticus strains, and that our PCR-DGGE analysis provided a more accurate picture of the population composition of whey starters compared to culture-dependent techniques that often fail to isolate the most abundant strains.
机译:由于该细菌种群在Grana Padano奶酪生产中所起的关键作用,因此在工业水平上监测天然乳清发酵剂中的瑞士乳杆菌菌株动力学十分重要。在这项研究中,我们旨在开发一种基于slpH基因座的PCR变性梯度凝胶电泳(PCR-DGGE)分析方法,同时使用优化的培养基进行乳清样品的培养依赖性分析,以使分离菌株的数量最大化。我们设计了针对slpH基因座的新引物,以扩增适合PCR-DGGE分析和区分菌株的基因区域。我们的结果证实,开发的PCR-DGGE方法可以快速,可靠地监测乳清发酵培养物中的瑞士乳杆菌种群。从乳清样品的PCR-DGGE图谱中检测到的所有条带序列均与瑞士伊豆的S层基因高度相似,并且与优势菌株的slpH基因座序列完全匹配。总体而言,我们的发现表明,slpH基因座的目标区域具有足够的异源性,可以区分瑞士乳杆菌。与通常依赖于培养物的技术相比,我们的PCR-DGGE分析提供了更准确的乳清发酵剂种群组成图。无法分离出最丰富的菌株。

著录项

  • 来源
    《Food microbiology》 |2020年第8期|103457.1-103457.6|共6页
  • 作者单位

    Centra Rkerche Biotecnologiche Universita Cattolca del Sacro Cuore Via Milano 24 26100 Cremona Italy;

    DiSTAS Department for Sustainable Food Process Universita Cattolica del Sacro Cuore Via E. Parmense 84 29122 Piacenza Italy;

    Centra Rkerche Biotecnologiche Universita Cattolca del Sacro Cuore Via Milano 24 26100 Cremona Italy DiSTAS Department for Sustainable Food Process Universita Cattolica del Sacro Cuore Via E. Parmense 84 29122 Piacenza Italy;

  • 收录信息 美国《科学引文索引》(SCI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    L. helveticus; PCR-DGGE; S-layer gene; Grana Padano; Natural whey cultures;

    机译:瑞士乳杆菌;PCR-DGGE;S层基因;Grana Padano;天然乳清文化;

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