Mining for Active Molecules in Probiotic Supernatant by Combining Non-Targeted Metabolomics and Immunoregulation Testing

Abstract

Chronic respiratory diseases such as asthma are highly prevalent in industrialized countries. As cases are expected to rise, there is a growing demand for alternative therapies. Our recent research on the potential benefits of probiotics suggests that they could prevent and reduce the symptoms of many diseases by modulating the host immune system with secreted metabolites. This article presents the first steps of the research that led us to identify the immunoregulatory bioactivity of the amino acid d-Trp reported in our previous study. Here we analyzed the cell culture metabolic footprinting of 25 commercially available probiotic strains to associate metabolic pathway activity information with their respective immune modulatory activity observed in vitro. Crude probiotic supernatant samples were processed in three different ways prior to untargeted analysis in positive and negative ionization mode by direct infusion ESI-FT-ICR-MS: protein precipitation and solid phase extraction (SPE) using HLB and CN-E sorbent cartridges. The data obtained were submitted to multivariate statistical analyses to distinguish supernatant samples into the bioactive and non-bioactive group. Pathway analysis using discriminant molecular features showed an overrepresentation of the tryptophan metabolic pathway for the bioactive supernatant class, suggesting that molecules taking part in that pathway may be involved in the immunomodulatory activity observed in vitro. This work showcases the potential of metabolomics to drive product development and novel bioactive compound discovery out of complex biological samples in a top-down manner.

Transient Mild Photothermia Improves Therapeutic Performance of Oral Nanomedicines with Enhanced Accumulation in the Colitis Mucosa

“…As reported, 3‐HAA possesses immune‐modulatory properties per se and strong antioxidant activities. [ 54 ] It can induce lymphocyte apoptosis and T cell suppression by inhibiting the NF‐ κ B signaling pathway, thereby decreasing the levels of proinflammatory cytokines (e.g., TNF‐ α and IL‐6). [ 55 ] Another study revealed that its anti‐inflammatory mechanism was associated with an increase in the level of Foxp3 + regulatory T cells.…”

Increased levels of a mycophenolic acid metabolite in patients with kidney failure negatively affect cardiomyocyte health

“…We previously described bioactivity-guided chromatography combined with ultrahigh-resolution mass spectrometry for the discovery of novel bioactive compounds ( 14 , 15 ). Here, the dried bioactive and inactive chromatographic fractions were solubilized in minimal volume of water (200 µl) diluted in methanol 1:5 (v:v) for direct injection at 2 µl/min into electrospray ionization negative mode ultrahigh-resolution mass spectrometry.…”

Effects of Probiotics on Gut Microbiota: An Overview

“…Gut microbial metabolites such as SCFAs, amino acids, bile acids, and vitamins involved in immune regulation are modulated by probiotics and influence the outcome of intestinal inflammation. SCFAs exert anti-inflammatory effects by binding to specific receptors on epithelial cells, inhibiting the production of pro-inflammatory cytokines [ 57 ]. Probiotic metabolites such as amino acids and amino acid derivatives interact with immune cell surface receptors and exert anti-inflammatory effects [ 57 ].…”

“…SCFAs exert anti-inflammatory effects by binding to specific receptors on epithelial cells, inhibiting the production of pro-inflammatory cytokines [ 57 ]. Probiotic metabolites such as amino acids and amino acid derivatives interact with immune cell surface receptors and exert anti-inflammatory effects [ 57 ]. An interesting effect of probiotics is their metabolization of primary bile acids to secondary bile acids which bind to various GPCR—G-protein coupled receptors or nuclear receptors to control mucosal immune activity and decrease inflammation [ 58 ].…”