Targeted Discovery of Glycosylated Natural Products by Tailoring Enzyme-Guided Genome Mining and MS-Based Metabolome Analysis

作者：Dawei Chen, Zhijun Song, Junjie Han, Jimei Liu, Hongwei Liu, Jungui Dai · 发表于：Journal of the American Chemical Society · 年份：2024 · DOI：10.1021/jacs.3c12895 · 被引用次数：25 · 研究领域：Microbial Natural Products and Biosynthesis、Fungal Biology and Applications、Plant biochemistry and biosynthesis

Glycosides make up a biomedically important class of secondary metabolites. Most naturally occurring glycosides were isolated from plants and bacteria; however, the chemical diversity of glycosylated natural products in fungi remains largely unexplored. Herein, we present a paradigm to specifically discover diverse and bioactive glycosylated natural products from fungi by combining tailoring enzyme-guided genome mining with mass spectrometry (MS)-based metabolome analysis. Through in vivo genes deletion and heterologous expression, the first fungal C -glycosyltransferase AuCGT involved in the biosynthesis of stromemycin was identified from Aspergillus ustus . Subsequent homology-based genome mining for fungal glycosyltransferases by using AuCGT as a probe revealed a variety of biosynthetic gene clusters (BGCs) containing its homologues in diverse fungi, of which the glycoside-producing capability was corroborated by high-performance liquid chromatography-mass spectrometry (HPLC-MS) analysis. Consequently, 28 fungal aromatic polyketide C / O -glycosides, including 20 new compounds, were efficiently discovered and isolated from the three selected fungi. Moreover, several novel fungal C / O -glycosyltransferases, especially three novel α-pyrone C -glycosyltransferases, were functionally characterized and verified in the biosynthesis of these glycosides. In addition, a proof of principle for combinatorial biosynthesis was applied to design the production of unnatural glycosides i...