Genomics-driven derivatization of the bioactive fungal sesterterpenoid variecolin: Creation of an unnatural analogue with improved anticancer properties

Yan, Dexiu; Arakelyan, Jemma; Wan, Teng; Raina, Ritvik; Chan, Tsz Ki; Ahn, Dohyun; Kushnarev, Vladimir; Cheung, Tsz Kiu; Chan, Ho Ching; Choi, Inseo; Ho, Pui Yi; Hu, Feijun; Kim, Yujeong; Lau, Hill Lam; Law, Ying Lo; Leung, Chi Seng; Tong, Chun Yin; Wong, Kai Kap; Yim, Wing Lam; Karnaukhov, Nikolay S.; Kong, Richard Y.C.; Babak, Maria V.; Matsuda, Yudai

doi:10.1016/j.apsb.2023.08.025

“…aculeatus ATCC 16872 that shares 65% amino acid sequence identity with EvAS has been reported to be responsible for the biosynthesis of variecoladiene ( 81 ) with a 5/8/6/5 fused ring system. 99 The authors speculated that VrcA shares carbocation intermediate I-5 + with EvAS, but triggers the C8 deprotonation of I-5 + to generate 81 .…”

Section: Terpenoid Cyclases In Fungal Terpenoid Biosynthesismentioning

confidence: 99%

“…During the period covered by this review, a detailed review of the post-tailoring enzymes associated with the biosynthesis of sesterterpenoid in fungi was published, with the exception of the newly identified oxidative enzyme in the biosynthesis of 81 (Scheme 37). 33,99…”

Section: Tailoring Enzymes In Fungal Terpenoid Biosynthesismentioning

confidence: 99%

“…179 Recently, Yan et al identified a biosynthetic gene cluster responsible for the biosynthesis of 252 and 253 in A. aculeatus ATCC 16872 using EvAS as a query sequence and elucidated the function of related genes through heterologous expression in A. oryzae NSAR1. 99 Sesterterpene cyclase VrcA catalyses the cyclization of GFPP to form the hydrocarbon skeleton 81 (Scheme 37), which is then oxidized by the P450 enzyme VrcB at sites C5 and C20 to form 252 , or followed by cyclization to produce 253 . In addition, the authors successfully identified four functional P450 enzymes by screening 17 other P450 enzymes from fungi that are involved or may be involved in the biosynthesis of terpenoids in combination with the cyclase VrcA.…”

Section: Tailoring Enzymes In Fungal Terpenoid Biosynthesismentioning

confidence: 99%

“…Remarkably, both 252 and 254 showed comparable anticancer activity in vitro and in vivo , with 254 showing lower toxic side effects in cancer-bearing mice. 99…”

Section: Tailoring Enzymes In Fungal Terpenoid Biosynthesismentioning

confidence: 99%

“…Accordingly, elucidation of the function of terpenoid cyclases and related tailoring enzymes will help to exploit the potential of these biocatalysts to discover second-generation terpenoids through pathway engineering. 19,99,137,178,188…”

Section: Conclusion and Prospectsmentioning

confidence: 99%

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Biosynthesis of fungal terpenoids

Luo,

Huang,

Lv

et al. 2024

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Function Switch of a Fungal Sesterterpene Synthase through Molecular Dynamics Simulation Assisted Alteration of an Aromatic Residue Cluster in the Active Pocket of PfNS

Zhang,

Wang,

Zhu

et al. 2024

Angewandte Chemie

0

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Terpene synthases (TPSs) play pivotal roles in generating diverse terpenoids through complex cyclization pathways. Protein engineering of TPSs offers a crucial approach to expanding terpene diversity. However, significant potential remains untapped due to limited understanding of the structure‐function relationships of TPSs. In this investigation, using a joint approach of molecular dynamics simulations‐assisted engineering and site‐directed mutagenesis, we manipulated the aromatic residue cluster (ARC) of a bifunctional terpene synthase (BFTPS), Pestalotiopsisfici nigtetraene synthase (PfNS). This led to the discovery of previously unreported catalytic functions yielding different cyclization patterns of sesterterpenes. Specifically, a quadruple variant (F89A/Y113F/W193L/T194W) completely altered PfNS's function, converting it from producing the bicyclic sesterterpene nigtetraene to the tricyclic ophiobolin F. Additionally, analysis of catalytic profiles by double, triple, and quadruple variants demonstrated that the ARC functions as a switch, unprecedently redirecting the production of 5/11 bicyclic (Type B) sesterterpenes to 5/15 bicyclic (Type A) ones. Molecular dynamics simulations and theozyme calculations further elucidated that, in addition to cation‐π interactions, C−H∙∙∙π interactions also play a key role in the cyclization patterns. This study offers a feasible strategy in protein engineering of TPSs for various industrial applications.

show abstract

Function Switch of a Fungal Sesterterpene Synthase through Molecular Dynamics Simulation Assisted Alteration of an Aromatic Residue Cluster in the Active Pocket of PfNS

Zhang,

Wang,

Zhu

et al. 2024

Angew Chem Int Ed

1

0

View full text Add to dashboard Cite

Terpene synthases (TPSs) play pivotal roles in generating diverse terpenoids through complex cyclization pathways. Protein engineering of TPSs offers a crucial approach to expanding terpene diversity. However, significant potential remains untapped due to limited understanding of the structure‐function relationships of TPSs. In this investigation, using a joint approach of molecular dynamics simulations‐assisted engineering and site‐directed mutagenesis, we manipulated the aromatic residue cluster (ARC) of a bifunctional terpene synthase (BFTPS), Pestalotiopsisfici nigtetraene synthase (PfNS). This led to the discovery of previously unreported catalytic functions yielding different cyclization patterns of sesterterpenes. Specifically, a quadruple variant (F89A/Y113F/W193L/T194W) completely altered PfNS's function, converting it from producing the bicyclic sesterterpene nigtetraene to the tricyclic ophiobolin F. Additionally, analysis of catalytic profiles by double, triple, and quadruple variants demonstrated that the ARC functions as a switch, unprecedently redirecting the production of 5/11 bicyclic (Type B) sesterterpenes to 5/15 bicyclic (Type A) ones. Molecular dynamics simulations and theozyme calculations further elucidated that, in addition to cation‐π interactions, C−H∙∙∙π interactions also play a key role in the cyclization patterns. This study offers a feasible strategy in protein engineering of TPSs for various industrial applications.

show abstract

Genomics-driven derivatization of the bioactive fungal sesterterpenoid variecolin: Creation of an unnatural analogue with improved anticancer properties

Cited by 5 publications

References 53 publications

Biosynthesis of fungal terpenoids

Biosynthesis of fungal terpenoids

Function Switch of a Fungal Sesterterpene Synthase through Molecular Dynamics Simulation Assisted Alteration of an Aromatic Residue Cluster in the Active Pocket of PfNS

Function Switch of a Fungal Sesterterpene Synthase through Molecular Dynamics Simulation Assisted Alteration of an Aromatic Residue Cluster in the Active Pocket of PfNS

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