Decoding Catalysis by Terpene Synthases

ACS Catalysis, Journal Year: 2023, Volume and Issue: 13(19), P. 12774 - 12802

Published: Sept. 15, 2023

The review by Christianson, published in 2017 on the twentieth anniversary of emergence field, summarizes foundational discoveries and key advances terpene synthase/cyclase (TS) biocatalysis (Christianson, D. W.

Language: Английский

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Computational and In silico study of novel fungicides against combating root rot, gray mold, fusarium wilt, and cereal rust

PLoS ONE, Journal Year: 2025, Volume and Issue: 20(1), P. e0316606 - e0316606

Published: Jan. 31, 2025

The exploration of potential candidates for fungicides against four fungal proteins that cause some vital plant diseases, namely Phytophthora capsici, Botrytis cinerea, Fusarium oxysporum f. sp. lycopersici, and Puccinia graminis tritici, was conducted using in silico, molecular docking simulations, dynamic (MD) simulation selecting the nature binding affinity with actives sites proteins. First all, DFT employed to optimize geometry, get prepared optimized ligand. From data, chemical descriptors were calculated. Next, two tools, such as AutoDock by PyRx Molecular Docking Glide from Schrödinger suite, used convey score, ligand protein interactions main proteases, instance 7VEM, 8H6Q, 8EBB, 7XDS having name pathogens: respectively. In case auto dock PyRx, L01, L03, L04, L13, L14, L17, L18, L19 demonstrated significantly higher affinities pathogens. Surprisingly, it is conveyed L03 illustrated highest score three L09 8H6Q. However, MD performed check validation calculation procedure stability docked complex accounting RMSD, RMSF, SASA, Radius gyration (Rg), Protein secondary structure elements (SSE), Ramachandran plot which confirm so high, number calculating hydrogen bonds more than good enough, a result concluded valid. Finally, Difenoconazole (L03) has been considered most promising antifungal drug evaluated studies.

Language: Английский

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Microbial‐type terpene synthases significantly contribute to the terpene profile of glandular trichomes of the fern Dryopteris fragrans (L.)

The Plant Journal, Journal Year: 2025, Volume and Issue: 121(6)

Published: March 1, 2025

SUMMARY Ferns, known for their adaptability and widespread presence, form a diverse group of plants. However, the mechanisms underlying terpenoid production, which are often linked to plant adaptation, not well understood in ferns. Here, we report that Dryopteris fragrans ( D. fragans ) produces terpenoids glandular trichomes (GTs) through activities microbial‐type terpene synthases. Using microscopy methods, capitate GTs were found occur on various organs throughout development . In leaves, 13 terpenoids, most being sesquiterpenoids, identified. By comparing chemistry intact GT‐removed isolated GTs, concluded be main site storage. Next, transcriptomes leaves created mined genes biosynthetic pathway. Among them nine putative full‐length microbial synthase‐like MTPSL designated DfMTPSL1–9 vitro enzyme assays, six DfMTPSLs demonstrated have sesquiterpene synthase activities. Of them, DfMTPSL1 catalyzes formation (−)‐9‐ epi ‐presilphiperfolan‐1‐ol, abundant sesquiterpenoid leaves. DfMTPS2 α‐muurolene, another major from The catalytic together with GT‐enriched expression respective support also biosynthesis Methyl jasmonate treatment induced DfMTPSL emission volatiles, suggesting GT‐produced play role defense against biotic stresses , similar counterparts seed

Language: Английский

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Engineered and total biosynthesis of fungal specialized metabolites

Nature Reviews Chemistry, Journal Year: 2024, Volume and Issue: 8(1), P. 61 - 78

Published: Jan. 3, 2024

Language: Английский

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Catalytic Mechanism and Heterologous Biosynthesis Application of Sesquiterpene Synthases

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(13), P. 6871 - 6888

Published: March 25, 2024

Sesquiterpenes comprise a diverse group of natural products with wide range applications in cosmetics, food, medicine, agriculture, and biofuels. Heterologous biosynthesis is increasingly employed for sesquiterpene production, aiming to overcome the limitations associated chemical synthesis extraction. Sesquiterpene synthases (STSs) play crucial role heterologous sesquiterpene. Under catalysis STSs, over 300 skeletons are produced through various cyclization processes (C1-C10 closure, C1-C11 C1-C6 C1-C7 closure), which responsible diversity sesquiterpenes. According types, we gave an overview advances understanding mechanism STSs from aspects protein crystal structures site-directed mutagenesis. We also summarized engineering Finally, bottlenecks potential research directions related application modified were presented.

Language: Английский

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Structural Insights Into the Terpene Cyclization Domains of Two Fungal Sesterterpene Synthases and Enzymatic Engineering for Sesterterpene Diversification

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(23)

Published: April 8, 2024

Abstract Little is known about the structures and catalytic mechanisms of sesterterpene synthases (StTSs), which greatly hinders structure‐based engineering StTSs for structural diversity expansion sesterterpenes. We here report on crystal terpene cyclization (TC) domains two fungal StTSs: sesterfisherol synthase (NfSS) sesterbrasiliatriene (PbSS). Both TC contain benzyltriethylammonium chloride (BTAC), pyrophosphate (PPi), magnesium ions (Mg 2+ ), clearly defining active sites. A combination theory experiments including carbocationic intermediates modeling, site‐directed mutagenesis, isotope labeling provided detailed insights into basis their mechanisms. Structure‐based NfSS PbSS resulted in formation 20 sesterterpenes 13 new compounds four pairs epimers with different configurations at C18. These results expand provide important future synthetic biology research.

Language: Английский

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Spiroluchuene A Synthase: A Cyclase from Aspergillus luchuensis Forming a Spirotetracyclic Diterpene

Angewandte Chemie International Edition, Journal Year: 2023, Volume and Issue: 62(52)

Published: Nov. 14, 2023

The diterpene synthase AlTS was identified from Aspergillus luchuensis. catalyses the formation of hydrocarbon spiroluchuene A, which exhibits a novel skeleton characterised by spirocyclic ring system. cyclisation mechanism towards this compound elucidated through isotopic labelling experiments in conjunction with DFT calculations and metadynamic simulations. biosynthetic intermediate luchudiene, besides derivative B, captured an enzyme variant obtained site-directed mutagenesis. With its 10-membered luchudiene is structurally related to germacrenes can undergo Cope rearrangement luchuelemene.

Language: Английский

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Sesquiterpene Cyclase BcBOT2 Promotes the Unprecedented Wagner-Meerwein Rearrangement of the Methoxy Group

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(26), P. 17838 - 17846

Published: June 18, 2024

Presilphiperfolan-8β-ol synthase (BcBOT2), a substrate-promiscuous sesquiterpene cyclase (STC) of fungal origin, is capable converting two new farnesyl pyrophosphate (FPP) derivatives modified at C7 bearing either hydroxymethyl group or methoxymethyl group. These substrates were chosen based on computationally generated model. Biotransformations yielded five oxygenated terpenoids. Remarkably, the formation one these tricyclic products can only be explained by cationically induced migration methoxy group, presumably via Meerwein-salt intermediate, unprecedented in synthetic chemistry and biosynthesis. The results show great principle general potential terpene cyclases for mechanistic studies unusual cation creation skeletons.

Language: Английский

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Computational Design-Enabled Divergent Modification of Monoterpene Synthases for Terpenoid Hyperproduction

ACS Catalysis, Journal Year: 2024, Volume and Issue: unknown, P. 17699 - 17715

Published: Nov. 15, 2024

Enzymes' catalytic promiscuity enables the alteration of product specificity via protein engineering; yet, harnessing this to achieve desired reactions remains challenging. Here, we identified HCinS, a monoterpene synthase (MTPS) with high efficiency and for 1,8-cineole biosynthesis. Quantum mechanics/molecular mechanics (QM/MM) simulations, which were performed based on resolved crystal structure revealed mechanistic details biosynthetic cascade reactions. Guided by these insights, in silico HCinS variants designed fine-tuned transition-state energies reaction microenvironments. Three (T111A, N135H, F236M), each one amino acid substitution, exhibited production monocyclic (R)-α-terpineol, (R)-limonene, acyclic myrcene, respectively, maintaining over 55% native HCinS. These surpassed naturally evolved isozymes capacity enabled yeast highest microbial titer corresponding terpene. Furthermore, single mutation four functional equivalent acids other TPSs, resulted expected shifts as variants. This research offers insights into mechanisms controlling TPS's highlights universal applicability computational design reshaping thereby paving innovative avenues creating enzymes applications chemistry synthetic biology.

Language: Английский

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Substrate-Dependent Mechanism Switch in the Desaturation Reactions of the Mononuclear Nonheme Iron Enzyme PtlD

ACS Catalysis, Journal Year: 2024, Volume and Issue: 14(10), P. 7389 - 7401

Published: April 27, 2024

PtlD, a multifunctional mononuclear nonheme iron and α-ketoglutarate-dependent (NHFe/α-KG) dioxygenase involved in neopentalenoketolactone biosynthesis, catalyzes hydroxylation, desaturation, olefin epoxidation reactions. Investigating desaturation reactions of nonactivated carbons mediated by NHFe/α-KG enzymes is intriguing, especially for understanding the fate substrate radicals formed after hydrogen atom abstraction FeIV═O species. Here, we investigate reaction mechanism PtlD using two distinct substrates: neopentalenolactone D (1) features lone pair-containing oxygen adjacent to olefin-forming carbon atoms, whereas pentalenolactone (7) harbors carbonyl group at corresponding position. For 1, our isotope effect measurement protein mutagenesis experiments suggest formation carbocation intermediate, which subsequently deprotonated base generate products. Residue K288 serves as base, while Y113 likely stabilizes via π-cation interaction. 7, incorporation patterns indicated that intermediate also but unstable, leading hydroxylation due H2O quenching. Notably, 7's exhibits temperature-dependent large kinetic (KIE) an inverse solvent (SIE), suggesting tunneling contributes electron–proton transfer (EPT) process. These findings collectively reveal cases enzymes, where mechanisms switch with different substrates.

Language: Английский

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