Abstract
The gut microbiome converts dietary compounds that are absorbed in the gastrointestinal tract and further metabolized by the human host. Sulfated metabolites are a major compound class derived from this co-metabolism and have been linked to disease development. In the present multidisciplinary study, we have investigated human urine samples from a dietary intervention study with 22 individuals collected before and after consumption of a polyphenol rich breakfast. These samples were analyzed utilizing our method combining enzymatic metabolite hydrolysis using an arylsulfatase and mass spectrometric metabolomics. Key to this study is the validation of 235 structurally diverse sulfated metabolites. We have identified 48 significantly upregulated metabolites upon dietary intervention including 11 previously unknown sulfated metabolites for this diet. We observed a large variation in subjects based on their potential to sulfate metabolites, which may be the foundation for classification of subjects as high and low sulfate metabolizers in future large cohort studies. The reported sulfatase-based method is a robust tool for the discovery of unknown microbiota-derived metabolites in human samples.
Highlights
Microbiota metabolism has been directly linked to human phy siology and disease development [1]
We sought to perform a detailed analysis of sulfated metabolites for large-scale identification of microbiota-host co-metabolism and dis covery of unknown bioactive metabolites in a two-tiered analysis
We employed our recently developed and efficient enzymatic assay for identification of sulfated metabolites in urine samples from different individuals, which requires enzymatic sample pretreatment followed by bioinformatic analysis for selective and sensitive identification of con verted sulfated metabolites using the metabolomics software package XCMS in R [24]
Summary
Microbiota metabolism has been directly linked to human phy siology and disease development [1]. Trillions of microbes reside in the human body carrying out biochemical conversions of metabolites or thogonal to the human hosts’ biochemical potential [2,3]. Metabolites produced through microbiome metabolism are known to have bioactive properties that can either be beneficial or toxic to the human body [4,5]. Single dietary compounds undergo several metabolic processes by diverse microbial species that result in a wide range of metabolites. Upon absorption of these microbe-derived metabolites, the human metabolic clearance mechanism further con verts these compounds prior to excretion from the body mainly through urine [8,9]. While several diet-specific metabolite classes have been identified, a plethora of metabolites produced after food consumption remains unknown as common analytical techniques are limited in their detection using mass spectrometric methods
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