Fungal indole alkaloid biogenesis through evolution of a bifunctional reductase/Diels–Alderase

Dan, Qingyun; Newmister, Sean A.; Klas, Kimberly R.; Fraley, Amy E.; McAfoos, Timothy J.; Somoza, Amber D.; Sunderhaus, James D.; Ye, Ying; Shende, Vikram V.; Yu, Fengan; Sanders, Jacob N.; Brown, W. Clay; Zhao, Le; Paton, Robert S.; Houk, K. N.; Smith, Janet L.; Sherman, David H.; Williams, Robert M.

doi:10.1038/s41557-019-0326-6

Title: Fungal indole alkaloid biogenesis through evolution of a bifunctional reductase/Diels–Alderase

Journal Article · Mon Sep 23 00:00:00 EDT 2019 · Nature Chemistry

DOI:https://doi.org/10.1038/s41557-019-0326-6· OSTI ID:1573393

^[1];

^[2]; Fraley, Amy E. ^[1]; McAfoos, Timothy J. ^[2]; Somoza, Amber D. ^[2]; Sunderhaus, James D. ^[2]; Ye, Ying ^[1]; Shende, Vikram V. ^[1]; Yu, Fengan ^[1]; Sanders, Jacob N. ^[3]; Brown, W. Clay ^[1]; Zhao, Le ^[2];

^[2]; Houk, K. N. ^[3];

^[1];

^[1]; Williams, Robert M. ^[2]

Univ. of Michigan, Ann Arbor, MI (United States)
Colorado State Univ., Fort Collins, CO (United States)
Univ. of California, Los Angeles, CA (United States)

Prenylated indole alkaloids such as the calmodulin-inhibitory malbrancheamides and anthelmintic paraherquamides possess great structural diversity and pharmaceutical utility. Here, we report complete elucidation of the malbrancheamide biosynthetic pathway accomplished through complementary approaches. These include a biomimetic total synthesis to access the natural alkaloid and biosynthetic intermediates in racemic form and in vitro enzymatic reconstitution to provide access to the natural antipode (+)-malbrancheamide. Reductive cleavage of an l-Pro–l-Trp dipeptide from the MalG non-ribosomal peptide synthetase (NRPS) followed by reverse prenylation and a cascade of post-NRPS reactions culminates in an intramolecular [4+2] hetero-Diels–Alder (IMDA) cyclization to furnish the bicyclo[2.2.2]diazaoctane scaffold. Enzymatic assembly of optically pure (+)-premalbrancheamide involves an unexpected zwitterionic intermediate where MalC catalyses enantioselective cycloaddition as a bifunctional NADPH-dependent reductase/Diels–Alderase. Furthermore, the crystal structures of substrate and product complexes together with site-directed mutagenesis and molecular dynamics simulations demonstrate how MalC and PhqE (its homologue from the paraherquamide pathway) catalyse diastereo- and enantioselective cyclization in the construction of this important class of secondary metabolites.

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Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: National Institutes of Health (NIH); National Science Foundation (NSF)

Grant/Contract Number:: R01 CA070375; R35 GM118101; R01 DK042303; F32GM122218; R01GM124480; OCI-1053575; AGM-12006; ACB-12002

OSTI ID:: 1573393

Journal Information:: Nature Chemistry, Vol. 11, Issue 11; ISSN 1755-4330

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 37 works

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37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
Biomimetic synthesis
biosynthesis
computational chemistry
enzyme mechanisms
x-ray crystallography

Title: Fungal indole alkaloid biogenesis through evolution of a bifunctional reductase/Diels–Alderase

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