Rapid differentiation of Lactobacillus species via metabolic profiling

Lactobacillus, the major genus of lactic acid bacteria group, plays functional roles in the human body, for example, convert sugars to lactic acid. They are the significant microbiota which can be found at a number of human body sites, such as the digestive system, urinary system, and genital system. A number of Lactobacillus species are often used as probiotics and can benefit host health when administered in adequate amounts. Due to their diverse functional characteristics, it is essential to have identification and high-resolution typing techniques to support the need in health and nutritional research of Lactobacillus species. In this study, we took advantages of both targeted and untargeted metabolomic technologies by using a triple quadrupole mass spectrometer (MS) in combination with a linear ion trap-Orbitrap hybrid MS, to investigate their capability and performance in deciphering the subtle metabolic difference in four closely related Lactobacillus species/strains. First, we evaluated the selected reaction monitoring (SRM) and high-resolution MS data for metabolite quantitation. Then the acquired data quality was further evaluated via the number of metabolites detected, the coefficient variation (CV) distribution, signal intensity distribution and so on. The established platforms were eventually applied to differentiate four Lactobacillus species in identical growth conditions. The proposed workflow demonstrated the capability of targeted and untargeted metabolomics in differentiating closely related bacterial strains/species.