We report the identification of the ter gene cluster responsible for the formation of the p-terphenyl derivatives terfestatins B and C and echoside B from the Appalachian Streptomyces strain RM-5-8. We characterize the function of TerB/C, catalysts that work together as a dual enzyme system in the biosynthesis of natural terphenyls. TerB acts as a reductase and TerC as a dehydratase to enable the conversion of polyporic acid to a terphenyl triol intermediate. X-ray crystallography of the apo and substrate-bound forms for both enzymes provides additional mechanistic insights. Validation of the TerC structural model via mutagenesis highlights a critical role of arginine 143 and aspartate 173 in catalysis. Cumulatively, this work highlights a set of enzymes acting in harmony to control and direct reactive intermediates and advances fundamental understanding of the previously unresolved early steps in terphenyl biosynthesis.
|Number of pages||9|
|Journal||ACS Chemical Biology|
|State||Published - Dec 17 2021|
Bibliographical noteFunding Information:
This work was supported by the Protein Structure Initiative NatPro Project (U01 GM098248), NIH grants R01 CA217255 (J.S.T. and G.N.P.) and R01 GM115261 (J.S.T. and G.N.P.), the Center of Biomedical Research Excellence (COBRE) in Pharmaceutical Research and Innovation (CPRI, NIH P20 GM130456), the University of Kentucky College of Pharmacy, and the National Center for Advancing Translational Sciences (UL1TR000117 and UL1TR001998). J.A.C. was also supported by the NIH NGMS Houston Area Molecular Biophysics Training Program (HAMBP, T32GM008280). This research also used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract no. DE-AC02-06CH11357. Use of LS-CAT Sector 21 was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (Grant 085P1000817). GM/CA@APS has been funded by the National Cancer Institute (ACB-12002) and the National Institute of General Medical Sciences (AGM-12006, P30GM138396). The Eiger 16M detector at GM/CA-XSD was funded by NIH grant S10 OD01228. In addition, we acknowledge the support of the University of Kentucky College of Pharmacy NMR Center and the Chapman University School of Pharmacy core laboratory facility.
© 2021 American Chemical Society
ASJC Scopus subject areas
- Molecular Medicine