Whole-cell-dependent biosynthesis of sulfo-conjugate using human sulfotransferase expressing budding yeast
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Cytosolic sulfotransferases (SULTs), one of the predominant phase II drug metabolizing enzymes (DME), play important roles in metabolism of xeno- and endobiotics to generate their sulfo-conjugates. These sulfo-conjugates often have biological activities but are difficult to study, because even though only small amounts are required to evaluate their efficacy and safety, chemical or biological synthesis of sulfo-conjugatesis is often challenging. Previously, we constructed a DME expression system for cytochrome P450 and UGT, using yeast cells, and successfully produced xenobiotic metabolites in a whole-cell-dependent manner. In this study, we developed a yeast expression system for human SULTs, including SULT1A1, 1A3, 1B1, 1C4, 1E1, and 2A1, in Saccharomyces cerevisiae and examined its sulfo-conjugate productivity. The recombinant yeast cells expressing each of the SULTs successfully produced several hundred milligram per liter of xeno- or endobioticsulfo-conjugates within 6 h. This whole-cell-dependent biosynthesis enabled us to produce sulfo-conjugates without the use of 3’-phosphoadenosine-5’-phosphosulfate, an expensive cofactor. Additionally, the production of regiospecific sulfo-conjugates of several polyphenols was possible with this method, making this novel yeast expression system a powerful tool for uncovering the metabolic pathways and biological actions of sulfo-conjugates.
KeywordsSulfotransferase (SULT) Sulfo-conjugate Regioselective biosynthesis Heterologous expression system in yeast Xenobiotic metabolism
The study was supported by the JSPSKAKENHI Grant Number JP26292072(SI).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Adamusová E, Cais O, Vyklický V, Kudová E, Chodounská H, Horák M, Vyklický L Jr (2013) Pregnenolone sulfate activates NMDA receptor channels. Physiol Res 62:731–736, 6Google Scholar
- Brand W, Boersma MG, Bik H, Hoek-van den Hil EF, Vervoort J, Barron D, Meinl W, Glatt H, Williamson G, van Bladeren PJ, Rietjens IM (2010) Phase II metabolism of hesperetin by individual UDP-glucuronosyltransferases and sulfotransferases and rat and human tissue samples. Drug Metab Dispos 38:617–625. https://doi.org/10.1124/dmd.109.031047 CrossRefPubMedGoogle Scholar
- Hehonah N, Zhu X, Brix L, Bolton-Grob R, Barnett A, Windmill K, McManus M (1999) Molecular cloning, expression, localisation and functional characterisation of a rabbit SULT1C2 sulfotransferase. Int J Biochem Cell Biol 31(8):869–882. https://doi.org/10.1016/S1357-2725(99)00038-2 CrossRefPubMedGoogle Scholar
- Ikushiro S, Nishikawa M, Masuyama Y, Shouji T, Fujii M, Hamada M, Nakajima N, Finel M, Yasuda K, Kamakura M, Sakaki T (2016) Biosynthesis of drug glucuronide metabolites in the budding yeast Saccharomyces cerevisiae. Mol Pharm 13:2274–2282. https://doi.org/10.1021/acs.molpharmaceut.5b00954 CrossRefPubMedGoogle Scholar
- Kasai N, Ikushiro S, Hirosue S, Arisawa A, Ichinose H, Wariishi H, Ohta M, Sakaki T (2009) Enzymatic properties of cytochrome P450 catalyzing 3′-hydroxylation of naringenin from the white-rot fungus Phanerochaete chrysosporium. Biochem Biophys Res Commun 387:103–108. https://doi.org/10.1016/j.bbrc.2009.06.134 CrossRefPubMedGoogle Scholar
- Kurata K, Takebayashi M, Morinobu S, Yamawaki S (2004) Beta-estradiol, dehydroepiandrosterone, and dehydroepiandrosterone sulfate protect against N-methyl-d-aspartate-induced neurotoxicity in rat hippocampal neurons by different mechanisms. J Pharmacol Exp Ther 311:237–245CrossRefPubMedGoogle Scholar
- LeblancN, WildeDW, KeefKD, HumeJR (1989) Electrophysiological mechanisms of minoxidil sulfate-induced vasodilation of rabbit portal vein. Circ Res65:1102–1011Google Scholar
- Patel KR, Andreadi C, Britton RG, Horner-Glister E, Karmokar A, Sale S, Brown VA, Brenner DE, Singh R, Steward WP, Gescher AJ, Brown K (2013) Sulfate metabolites provide an intracellular pool for resveratrol generation and induce autophagy with senescence. Sci Transl Med 5(205):205ra133. https://doi.org/10.1126/scitranslmed.3005870 CrossRefPubMedGoogle Scholar
- Sakaki T, Oeda K, Miyoshi M, Ohkawa H (1985) Characterization of rat cytochrome P-450MC synthesized in Saccharomyces cerevisiae. J Biochem, 98:167–175Google Scholar
- Sakakibara Y, Yanagisawa K, Katafuchi J, Ringer DP, Takami Y, Nakayama T, Suiko M, Liu MC (1998) Molecular cloning, expression, and characterization of novel human SULT1C sulfotransferases that catalyze the sulfonation of N-hydroxy-2-acetylaminofluorene. J Biol Chem 273:33929–33935CrossRefPubMedGoogle Scholar
- Shimohira T, Kurogi K, Hashiguchi T, Liu MC, Suiko M, Sakakibara Y (2017) Regioselective production of sulfated polyphenols using human cytosolic sulfotransferase-expressing Escherichia coli cells. J BiosciBioengpii S1389-1723(16):30404–30402. https://doi.org/10.1016/j.jbiosc.2017.02.006 Google Scholar
- Walker J, Schueller K, Schaefer LM, Pignitter M, Esefelder L, Somoza V (2014) Resveratrol and its metabolites inhibit pro-inflammatory effects of lipopolysaccharides in U-937 macrophages in plasma-representative concentrations. Food Funct 5:74–84. https://doi.org/10.1039/c3fo60236b CrossRefPubMedGoogle Scholar
- Zhang M, Jagdmann GE Jr, Van Zandt M, Sheeler R, Beckett P, Schroeter H (2013) Chemical synthesis and characterization of epicatechin glucuronides and sulfates: bioanalytical standards for epicatechin metabolite identification. J Nat Prod 76:157–169. https://doi.org/10.1021/np300568m CrossRefPubMedGoogle Scholar