Total biosynthesis or whole-cell biocatalytic production of sulfated small molecules relies on the discovery and implementation of appropriate sulfotransferase enzymes. Although fungi are prominent biocatalysts and have been used to sulfate drug-like phenolics, no gene encoding a sulfotransferase enzyme has been functionally characterized from these organisms. Here, we identify a phenolic sulfotransferase, FgSULT1, by genome mining from the plant-pathogenic fungus Fusarium graminearum PH-1. We expressed FgSULT1 in a Saccharomyces cerevisiae chassis to modify a broad range of benzenediol lactones and their nonmacrocyclic congeners, together with an anthraquinone, with the resulting unnatural natural product (uNP) sulfates displaying increased solubility. FgSULT1 shares low similarity with known animal and plant sulfotransferases. Instead, it forms a sulfotransferase family with putative bacterial and fungal enzymes for phase II detoxification of xenobiotics and allelochemicals. Among fungi, putative FgSULT1 homologues are encoded in the genomes of Fusarium spp. and a few other genera in nonsyntenic regions, some of which may be related to catabolic sulfur recycling. Computational structure modeling combined with site-directed mutagenesis revealed that FgSULT1 retains the key catalytic residues and the typical fold of characterized animal and plant sulfotransferases. Our work opens the way for the discovery of hitherto unknown fungal sulfotransferases and provides a synthetic biological and enzymatic platform that can be adapted to produce bioactive sulfates, together with sulfate ester standards and probes for masked mycotoxins, precarcinogenic toxins, and xenobiotics.

中文翻译：

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硫酸化苯二醇内酯与来自禾谷镰刀菌 PH-1 的酚类磺基转移酶的组合生物合成


硫酸化小分子的总生物合成或全细胞生物催化生产依赖于适当的磺基转移酶的发现和实施。尽管真菌是重要的生物催化剂并且已被用于硫酸化类药物酚类物质，但尚未从这些生物体中对编码磺基转移酶的基因进行功能表征。在这里，我们通过植物病原真菌禾谷镰刀菌PH-1 的基因组挖掘鉴定出一种酚类磺基转移酶 FgSULT1。我们在酿酒酵母底盘中表达 FgSULT1，以修饰多种苯二酚内酯及其非大环同系物以及蒽醌，所得非天然天然产物 (uNP) 硫酸盐显示出更高的溶解度。 FgSULT1 与已知的动植物磺基转移酶相似度较低。相反，它与假定的细菌和真菌酶形成磺基转移酶家族，用于异生物质和化感物质的 II 期解毒。在真菌中，假定的 FgSULT1 同源物编码在镰刀菌属 ( Fusarium spp) 的基因组中。以及非同线区域的其他一些属，其中一些可能与分解代谢硫回收有关。计算结构建模与定点诱变相结合表明，FgSULT1 保留了关键催化残基和特征性动植物磺基转移酶的典型折叠。我们的工作为发现迄今为止未知的真菌磺基转移酶开辟了道路，并提供了一个合成的生物和酶平台，可用于生产生物活性硫酸盐，以及硫酸酯标准品和掩盖霉菌毒素、致癌前毒素和异生物质的探针。