Institute of Microbiology of the Czech Academy of Sciences, Vídeňská 1083, Prague 4 CZ-142 00, Czech Republic.
Department of Biochemistry and Microbiology, University of Chemistry and Technology Prague, Technická 3, Prague 6 CZ-166 28, Czech Republic.
J Agric Food Chem. 2024 Oct 9;72(40):22208-22216. doi: 10.1021/acs.jafc.4c06771. Epub 2024 Oct 1.
The preparation of pure metabolites of bioactive compounds, particularly (poly)phenols, is essential for the accurate determination of their pharmacological profiles . Since the extraction of these metabolites from biological material is tedious and impractical, they can be synthesized enzymatically by bacterial PAPS-independent aryl sulfotransferases (ASTs). However, only a few ASTs have been studied and used for (poly)phenol sulfation. This study introduces new fully characterized recombinant ASTs selected according to their similarity to the previously characterized ASTs. These enzymes, produced in , were purified, biochemically characterized, and screened for the sulfation of nine flavonoids and two phenolic acids using nitrophenyl sulfate. All tested compounds were proved to be substrates for the new ASTs, with kaempferol and luteolin being the best converted acceptors. ASTs from (AST) and (AST) showed the highest efficiency in the sulfation of tested polyphenols. To demonstrate the efficiency of the present sulfation approach, a series of new authentic metabolite standards, regioisomers of kaempferol sulfate, were enzymatically produced, isolated, and structurally characterized.
生物活性化合物(尤其是(多)酚类化合物)的纯代谢物的制备对于准确确定其药理特性至关重要。由于从生物材料中提取这些代谢物既繁琐又不切实际,因此可以通过细菌 PAPS 非依赖性芳基磺基转移酶(AST)酶促合成。然而,只有少数 AST 已被研究并用于(多)酚的磺化。本研究根据与先前表征的 AST 的相似性,引入了新的经过充分表征的重组 AST。这些在 中生产的酶经过纯化、生物化学表征,并使用对硝基苯硫酸盐筛选了 9 种类黄酮和 2 种酚酸的磺化反应。所有测试的化合物均被证明是新 AST 的底物,其中山柰酚和木犀草素是最佳的转化受体。来自 (AST)和 (AST)的 AST 在测试多酚的磺化中表现出最高的效率。为了证明本磺化方法的效率,使用酶法产生、分离和结构表征了一系列新的真实代谢物标准品,即山柰酚硫酸盐的区域异构体。