Biological and Medicinal Chemistry

Discovery, structure, and mechanism of a class II sesquiterpene cyclase

Authors

  • Xingming Pan State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, Jiangsu, China ,
  • Wenyu Du State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, Jiangsu, China ,
  • Xiaowei Zhang State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, Jiangsu, China ,
  • Xiaoxu Lin State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, Jiangsu, China ,
  • Fang-Ru Li State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, Jiangsu, China ,
  • Qian Yang State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, Jiangsu, China ,
  • Hang Wang State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, Jiangsu, China ,
  • Jeffrey D. Rudolf Department of Chemistry, University of Florida, Gainesville, Florida 32611-7011, United States ,
  • Bo Zhang State Key Laboratory of Pharmaceutical Biotechnology, Institute of Functional Biomolecules, School of Life Sciences, Nanjing University, Nanjing 210023, Jiangsu, China ,
  • Liao-Bin Dong State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, Jiangsu, China

Abstract

Terpene cyclases (TCs), the extraordinary enzymes that create the structural diversity seen in terpene natural products, are traditionally divided into two classes. Although the structural and mechanistic features in class I TCs are well-known, the corresponding details in class II counterparts have not been fully characterized. Here, we report the genome mining discovery and structural characterization of two class II sesquiterpene cyclases (STCs) from Streptomyces. These drimenyl diphosphate synthases (DMSs) are the first STCs shown to possess β,γ-didomain architecture. High-resolution X-ray crystal structures of SsDMS in complex with both a farnesyl diphosphate and Mg2+ unveiled an induced-fit mechanism with an unprecedented Mg2+ binding mode, finally solving one of the lingering questions in class II TC enzymology. This study supports continued genome mining for novel bacterial TCs and provides new mechanistic insights into canonical class II TCs that will lead to advances in TC engineering and synthetic biology.

Content

Thumbnail image of SsDMS_Manuscript 082422f.pdf

Supplementary material

Thumbnail image of SsDMS_SI_082322f.pdf
Supplementary Information for Discovery, structure and mechanism of a class II sesquiterpene cyclase
The Supporting Information includes materials, methods, detailed experimental procedures, bioinformatic analysis, in vivo, in vitro, and structural characterizations of SsDMS.