Gut microbes have been recognized to convert human bile acids by deconjugation, dehydroxylation, dehydrogenation, and epimerization of the cholesterol core, but the ability to re-conjugate them with amino acids as an additional conversion has been recently described. These new bile acids are known as microbially conjugated bile acids (MCBAs). The aim of this study was to evaluate the MCBAs diversity produced by the gut microbiota through a metabolomics approach. In this study, fresh fecal samples from healthy donors were evaluated to explore the re-conjugation of chenodeoxycholic and 3-oxo-chenodeoxycholic acids by the human gut microbiota. No significant differences were found between the conversion trend of both BAs incubations. The in vitro results showed a clear trend to first accumulate the epimer isoursochenodeoxycholic acid and the dehydroxylated lithocholic acid derivatives in samples incubated with chenodeoxycholic and 3-oxo-chenodeoxycholic acid. They also showed a strong trend for the production of microbially conjugated dehydroxylated bile acids instead of chenodeoxycholic backbone conjugates. Different molecules and isomers of MCBAs were identified, and the new ones, valolithocholate ester and leucolithocholate ester, were identified and confirmed by MS/MS. These results document the gut microbiota’s capability to produce esters of MCBAs on hydroxyls of the sterol backbone in addition to amides at the C24 acyl site. This study opens a new perspective to study the BAs diversity produced by the human gut microbiota.

The control and development of plant growth promoters is a key factor for the agro-nomy industry in its economic performance. Different genera of bacteria are widely used as natural biostimulants with the aim of enhancing nutrition efficiency, abiotic stress tolerance and/or crop quality traits, regardless of their nutrients content. However, the complete exo-metabolome of the bacteria responsible for the biostimulant effect is still unknown and needs to be investigated. Three bacteria with different biostimulant effects were studied by untargeted metabolomics in order to describe the metabolites responsible for this effect. The pentose phosphate pathway, tryptophan metabolism, zeatin biosynthesis, vitamin B6 metabolism and amino acid metabolism were the highlighted pathways related to bacteria biostimulant activity. These results are related to the plant hormones biosynthesis pathway for auxins and zeatins biosynthesis. Fourteen metabolites were identified as biomarkers of the biostimulant activity. The results suggest a greater relevance of auxins than cytokinin pathways due the importance of the precursors identified. The results show a clear trend of using indole-3-pyruvate and 3-Indoleglycolaldehyde pathways to produce auxins by bacteria. The results demonstrate for the first time that 4-Pyridoxic acid, the fructosamines N-(1-Deoxy-1-fructosyl)phenylalanine and N-(1-Deoxy-1-fructosyl)isoleucine and the tripeptides diprotin A and B are metabolites related to biostimulant capabilities. This study shows how untargeted metabolomic approaches can be useful tools to investigate the bacteria exo-metabolomes related to biostimulant effects.