Regioselective production of sulfated polyphenols using human cytosolic sulfotransferase-expressing Escherichia coli cells

Abstract

Dietary polyphenols present in fruits and vegetables have been reported to manifest beneficial health effects on humans. Polyphenol metabolites including their sulfated derivatives have been shown to be biologically active. Primarily due to the difficulty in preparing regiospecific sulfated polyphenols for detailed investigations, the exact functions of sulfated polyphenols, however, remain unclear. The current study aimed to develop a procedure for the regioselective production of sulfated polyphenols using E. coli cells expressing human cytosolic sulfotransferases (SULTs). Two regioisomers of sulfated genistein were produced by E. coli cells expressing human SULT1A3, SULT1C4, or SULT1E1, and purified using Diaion HP20 resin, followed by high pressure liquid chromatography (HPLC). Structural analysis using mass spectrometry (MS) and nuclear magnetic resonance (NMR) revealed that E. coli cells expressing SULT1A3 preferentially produced genistein 4’-sulfate, whereas E. coli cells expressing SULT1C4 preferentially produced genistein 7-sulfate. To improve the bioproductivity, the effects of several factors including the concentrations of glucose and SO42−, and growth temperature were investigated. The bioproduction procedure established in this study will be valuable for the production of regioselective sulfated polyphenols for use in future studies on their biological functions.

Novel silkworm (Bombyx mori) sulfotransferase swSULT ST3 is involved in metabolism of polyphenols from mulberry leaves

“…This suggests that the metabolite was mono-sulfated genistein. Similar observation was reported for human SULT1C4 which catalyzed the production of genistein 7-sulfate [ 33 ].…”

“…This suggests that the metabolite was mono-sulfated genistein. Similar observation was reported for human SULT1C4 which catalyzed the production of genistein 7-sulfate [33]. Similar to swSULT ST1 and ST2 (Table 1), swSULT ST3 can catalyze xanthurenic acid.…”

From Steroid and Drug Metabolism to Glycobiology, Using Sulfotransferase Structures to Understand and Tailor Function

“…Particularly, a triple mutation of zf6-OST-3 designed based on the crystal structures, allows for sulfation only on the non-reducing terminal glucosamine, providing fine-tuned control of 6-O-sulfation in the chemoenzymatic synthesis pathway (Yi et al, 2020). Cytosolic sulfotransferases are also being investigated for their ability to generate sulfated products for biological reagents and drug discovery (Lamb et al, 2006;Matsumura et al, 2018;Shi et al, 2012;Shimohira et al, 2017;Xie et al, 2020).…”

Metabolic Engineering of Escherichia coli for Enhanced Production of Naringenin 7-Sulfate and Its Biological Activities

“…Enzymatic sulfation is catalyzed by a family of sulfotransferases (STs) that transfer the sulfonate group of 3′-phosphoadenosine-5′-phosphosulfate (PAPS) to a hydroxyl group or amino group of acceptor compounds with the parallel formation of 3′-phosphoadenosine-5′-phosphate (PAP) ( Paul et al, 2012 ). Although the sequences of many STs have been deposited to various genome databases of plant ( Gidda and Varin, 2006 ; Marsolais et al, 2007 ), bacteria ( Hossain et al, 2011 ), and mammalian ( Wong et al, 2010 ; Shimohira et al, 2017 ), the enzymatic and microbial synthesis of sulfonated natural products still are limited. One of the major problems of in vitro sulfation reactions is the use of the costly and unstable sulfate donor, PAPS, which impedes industrial scale-up of the process.…”