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Development of omics‐based protocols for the microbiological characterization of multi‐strain formulations marketed as probiotics: the case of VSL#3

机译:基于OMICS的微生物学表征的遗传多重制剂作为益生菌的微生物特征的发展:VSL#3的情况

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摘要

Summary The growing commercial interest in multi‐strain formulations marketed as probiotics has not been accompanied by an equal increase in the evaluation of quality levels of these biotechnological products. The multi‐strain product VSL#3 was used as a model to setup a microbiological characterization that could be extended to other formulations with high complexity. Shotgun metagenomics by deep Illumina sequencing was applied to DNA isolated from the commercial VSL#3 product to confirm strains identity safety and composition. Single‐cell analysis was used to evaluate the cell viability, and β‐galactosidase and urease activity have been used as marker to monitor the reproducibility of the production process. Similarly, these lots were characterized in detail by a metaproteomics approach for which a robust protein extraction protocol was combined with advanced mass spectrometry. The results identified over 1600 protein groups belonging to all strains present in the VSL#3 formulation. Of interest, only 3.2 % proteins showed significant differences mainly related to small variations in strain abundance. The protocols developed in this study addressed several quality criteria that are relevant for marketed multi‐strain products and these represent the first efforts to define the quality of complex probiotic formulations such as VSL#3.
机译:发明内容由于益生菌销售的多重应变配方的日益增长的商业兴趣尚未伴随着这些生物技术产品质量水平评估的同等增加。多应变产物VSL#3用作设置微生物表征的模型,该表征可以扩展到具有高复杂性的其他配方。通过Deep Illumina测序的霰弹枪Metagenomics应用于从商业VSL#3产品分离的DNA,以确认菌株标识安全性和组成。单细胞分析用于评估细胞活力,β-半乳糖苷酶和脲酶活性已被用作标记以监测生产过程的再现性。类似地,这些批次通过稳健的蛋白质提取方案与晚期质谱结合的元标瘤方法详细描述了这些批次。结果鉴定了属于VSL#3制剂中存在的所有菌株的1600多种蛋白质基团。感兴趣的是,只有3.2%的蛋白质显示出显着差异,主要是与应变丰度的小变化有关。本研究中开发的协议解决了几种质量标准,该质量标准与销售的多重品产有关,这些质量标准代表了定义复合益生菌制剂质量如VSL#3的第一努力。

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