PDF(Pubmed)
Abstract：
Probiotic bacteria, capable of conferring benefits to the host, can present challenges in design, development, scale-up, manufacturing, commercialization, and life cycle management. Strain identification is one of the main quality parameters; nevertheless, this task can be challenging since established methodologies can lack resolution at the strain level for some microorganisms and\\or are labor-intensive and time-consuming. Fourier transform infrared spectroscopy (FTIRS) has been largely used for the investigation of pathogenic species in the clinical field, whereas only recently has been proposed for the identification of probiotic strains. Within the probiotic industrial production, bacterial strains can be subjected to stressful conditions that may affect genomic and phenotypic characteristics; therefore, real-time monitoring of all the sequential growth steps is requested. Considering the fast, low-cost, and high-throughput features, FTIRS is an innovative and functional technology for typing probiotic strains from bench-top experiments to large-scale industrial production, allowing the monitoring of stability and identity of probiotic strains. In this study, the discriminatory power of FTIRS was assessed for four Lactiplantibacillus plantarum probiotic strains grown under different conditions, including temperatures (30 and 37°C) and medium (broth and agar), after consecutive sub-culturing steps. A comparison between the generated spectra with pulsed-field gel electrophoresis (PFGE) profiles was also performed. FTIRS was not only able to distinguish the strains of L. plantarum under different growth conditions but also to prove the phenotypic stability of L. plantarum type strain LP-CT after six growing steps. Regardless of the growth conditions, FTIRS spectra related to LP-CT constituted a unique hierarchical cluster, separated from the other L. plantarum strains. These results were confirmed by a PFGE analysis. In addition, based on FTIRS data, broth cultures demonstrated a higher reproducibility and discriminatory power with respect to agar ones. These results support the introduction of FTIRS in the probiotic industry, allowing for the step-by-step monitoring of massive microbial production while also guaranteeing the stability and purity of the probiotic strain. The proposed novel approach can constitute an impressive improvement in the probiotic manufacturing process.
摘要：
益生菌,能够给主人带来好处，可以在设计中提出挑战，发展,扩大规模，制造,商业化，和生命周期管理。应变识别是主要的质量参数之一；然而，这项任务可能具有挑战性，因为已建立的方法可能缺乏某些微生物在菌株水平上的分辨率，并且\\或者是劳动密集型和耗时的。傅里叶变换红外光谱（FTIRS）已被广泛用于临床领域的病原物种的研究，而直到最近才被提议用于鉴定益生菌菌株。在益生菌工业生产中，细菌菌株可以经受可能影响基因组和表型特征的应激条件;因此,要求对所有连续生长步骤进行实时监控。考虑到快,低成本,和高通量功能，FTIRS是一项创新和功能技术，用于从台式实验到大规模工业生产的益生菌菌株分型，允许监测益生菌菌株的稳定性和身份。在这项研究中,对在不同条件下生长的四种植物乳杆菌益生菌菌株进行了FTIRS的辨别能力评估，包括温度（30和37°C）和培养基（肉汤和琼脂），在连续的传代培养步骤后。还进行了所产生的光谱与脉冲场凝胶电泳（PFGE）曲线之间的比较。FTIRS不仅能够区分不同生长条件下的植物乳杆菌菌株，而且能够证明植物乳杆菌型菌株LP-CT经过六个生长步骤后的表型稳定性。不管生长条件如何，与LP-CT相关的FTIRS光谱构成了一个独特的分层簇，与其他植物乳杆菌菌株分离。这些结果通过PFGE分析得到证实。此外,基于FTIRS数据，肉汤培养物相对于琼脂培养物表现出更高的可重复性和辨别能力。这些结果支持FTIRS在益生菌行业的引入，允许逐步监测大规模微生物生产，同时保证益生菌菌株的稳定性和纯度。所提出的新方法可以构成益生菌制造过程中令人印象深刻的改进。
