Shimohira Takehiko, Kurogi Katsuhisa, Hashiguchi Takuyu, Liu Ming-Cheh, Suiko Masahito, Sakakibara Yoichi
Department of Biochemistry and Applied Biosciences, University of Miyazaki, 1-1 Gakuenkibanadai-Nishi, Miyazaki 889-2192, Japan; Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki, 1-1 Gakuenkibanadai-Nishi, Miyazaki 889-2192, Japan.
Department of Pharmacology, College of Pharmacy and Pharmaceutical Sciences, University of Toledo, Toledo, OH 43614, USA.
J Biosci Bioeng. 2017 Jul;124(1):84-90. doi: 10.1016/j.jbiosc.2017.02.006. Epub 2017 Mar 9.
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 Escherichia 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 SO, 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.
据报道，水果和蔬菜中的膳食多酚对人体具有有益的健康影响。包括其硫酸化衍生物在内的多酚代谢产物已被证明具有生物活性。然而，主要由于难以制备区域特异性硫酸化多酚用于详细研究，硫酸化多酚的确切功能仍不清楚。本研究旨在开发一种使用表达人胞质硫酸转移酶（SULTs）的大肠杆菌细胞区域选择性生产硫酸化多酚的方法。表达人SULT1A3、SULT1C4或SULT1E1的大肠杆菌细胞产生了硫酸化染料木黄酮的两种区域异构体，并使用Diaion HP20树脂进行纯化，然后进行高压液相色谱（HPLC）。使用质谱（MS）和核磁共振（NMR）进行的结构分析表明，表达SULT1A3的大肠杆菌细胞优先产生染料木黄酮4'-硫酸盐，而表达SULT1C4的大肠杆菌细胞优先产生染料木黄酮7-硫酸盐。为了提高生物生产力，研究了包括葡萄糖和SO浓度以及生长温度在内的几个因素的影响。本研究建立的生物生产方法对于生产区域选择性硫酸化多酚用于其生物学功能的未来研究将具有重要价值。
