Comparison of two arylsulfatases for targeted mass spectrometric analysis of microbiota-derived metabolites

Sulfation of metabolites is the second highest phase II modification in humans, which plays a critical role xenobiotics clearance process and gut microbiota-host co-metabolism. Besides main function to remove from body, sulfated have also been linked inflammation, bacterial pathogenesis metabolic disorders. A better understanding how these impact human body has turned into an important research area. Analytical methods for selective identification this metabolite class are scarce. We recently developed assay utilizing arylsulfatase Helix pomatia due high substrate promiscuity combined with state-of-the-art metabolomics bioinformatic analysis O-sulfated samples. This enzyme requires multistep purification as purity needed mass spectrometric assay. In study, we utilized new recombinant overexpressed (ASPC) organic sulfate esters urine compared conversion samples specificity purified pomatia. Our revealed that both enzymes can be discovery demonstrated by equal hydrolysis 108 substrates including conjugates 27 microbial origin. Importantly, identified 21 exclusively hydrolyzed ASPC application increases unknown higher scaffold diversity.