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Comparison of two a...
Comparison of two arylsulfatases for targeted mass spectrometric analysis of microbiota-derived metabolites
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- Jain, Abhishek (author)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Institutionen för kemi - BMC
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- Correia, Mario S. P. (author)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Analytisk kemi
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- Meistermann, Hannes (author)
- Uppsala universitet,Institutionen för kemi - BMC,Science for Life Laboratory, SciLifeLab
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- Vujasinovic, Miroslav (author)
- Karolinska Univ Hosp, Dept Digest Dis, Stockholm, Sweden.
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- Lohr, J-Matthias (author)
- Karolinska Institutet,Karolinska Univ Hosp, Dept Digest Dis, Stockholm, Sweden.;Karolinska Inst, Dept Clin Sci Intervent & Technol CLINTEC, Stockholm, Sweden.
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- Globisch, Daniel (author)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Analytisk kemi
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(creator_code:org_t)
- Elsevier, 2021
- 2021
- English.
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In: Journal of Pharmaceutical and Biomedical Analysis. - : Elsevier. - 0731-7085 .- 1873-264X. ; 195
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Abstract
Subject headings
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- Sulfation of metabolites is the second highest phase II modification in humans, which plays a critical role in the xenobiotics clearance process and gut microbiota-host co-metabolism. Besides the main function to remove xenobiotics from the body, sulfated metabolites have also been linked to inflammation, bacterial pathogenesis and metabolic disorders. A better understanding of how these metabolites impact the human body has turned into an important research area. Analytical methods for selective identification of this metabolite class are scarce. We have recently developed an assay utilizing the arylsulfatase from Helix pomatia due to a high substrate promiscuity combined with state-of-the-art metabolomics bioinformatic analysis for the selective identification of O-sulfated metabolites in human samples. This enzyme requires a multistep purification process as highest purity is needed for the developed mass spectrometric assay. In this study, we have utilized a new and recombinant overexpressed arylsulfatase (ASPC) for the selective identification of organic sulfate esters in human urine samples. We have compared the substrate conversion in urine samples and substrate specificity of this enzyme with purified arylsulfatase from Helix pomatia. Our analysis of urine samples revealed that both enzymes can be utilized for the selective analysis and discovery of sulfated metabolites with high promiscuity as demonstrated by equal hydrolysis of 108 substrates including sulfated conjugates of 27 metabolites of microbial origin. Importantly, we also identified 21 substrates in human urine samples that are exclusively hydrolyzed by ASPC and application of this enzyme increases the discovery of unknown sulfated metabolites with a higher scaffold diversity.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Keyword
- Arylsulfatase
- Mass spectrometry
- Microbiome
- Xenobiotics
- Enzyme activity
- Sulfated metabolites
Publication and Content Type
- ref (subject category)
- art (subject category)
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