Details
| Original language | English |
|---|---|
| Pages (from-to) | 8125-8139 |
| Number of pages | 15 |
| Journal | ACS catalysis |
| Volume | 15 |
| Issue number | 10 |
| Early online date | 1 May 2025 |
| Publication status | Published - 16 May 2025 |
Abstract
The fungal sesquiterpene synthase BcBOT2 shows unique substrate promiscuity. It transforms farnesyl pyrophosphate (FPP) into presilphiperfolan-8β-ol via a cationic cascade that is initiated by a (1 → 11) cyclization. Here, it is shown that BcBOT2 also accepts (2,3-Z)-configured FPP derivatives, which provide terpenoids that result from an initial (1 → 6) cyclization. “Methyl mapping” was conducted by shifting the position of one or more methyl groups, and it was found that the location of methyl groups has a profound effect on the efficacy of cyclizations. In particular, the shift of the methyl group at C3 to the C2 position has the most profound effect on cyclohexane formation. Molecular modeling studies show that (1 → 6) cyclization took place due to adopting a different catalytically competent docking pose of FPP derivatives compared to natural FPP within the active site of the BcBOT2, which is mainly due to the (2,3-Z)-configuration. This docking pose leads to a C1-C6 distance shorter (3.3-3.5 Å) than the typical association with a near-attack conformation required for cyclization. Finally, these biotransformation results were benchmarked by a comprehensive study of the “hydrolysis” of eight different FPP derivatives. Under enzyme-free conditions, cyclohexene and cycloheptene terpenoids are formed. The ring size is mainly determined by the position of the methyl group at C6 or C7. The study reveals that in addition to protein engineering, unnatural substrates can also be used to specifically manipulate the mode of cyclization of terpene synthases.
Keywords
- biotransformations, computational modeling, sesquiterpene, sesquiterpene synthases, terpenoids
ASJC Scopus subject areas
- Chemical Engineering(all)
- Catalysis
- Chemistry(all)
- General Chemistry
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In: ACS catalysis, Vol. 15, No. 10, 16.05.2025, p. 8125-8139.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Reprogramming the Cyclization of the Sesquiterpene Synthase BcBOT2 Using 2,3-Z-Configured FPP Derivatives and by Means of “Methyl Mapping”
AU - Budde, Jan Luca
AU - Çay, Muhammed Yasin
AU - Dräger, Gerald
AU - Droste, Jörn
AU - Hassanin, Ahmed
AU - Davari, Mehdi D.
AU - Kirschning, Andreas
N1 - Publisher Copyright: © 2025 The Authors. Published by American Chemical Society.
PY - 2025/5/16
Y1 - 2025/5/16
N2 - The fungal sesquiterpene synthase BcBOT2 shows unique substrate promiscuity. It transforms farnesyl pyrophosphate (FPP) into presilphiperfolan-8β-ol via a cationic cascade that is initiated by a (1 → 11) cyclization. Here, it is shown that BcBOT2 also accepts (2,3-Z)-configured FPP derivatives, which provide terpenoids that result from an initial (1 → 6) cyclization. “Methyl mapping” was conducted by shifting the position of one or more methyl groups, and it was found that the location of methyl groups has a profound effect on the efficacy of cyclizations. In particular, the shift of the methyl group at C3 to the C2 position has the most profound effect on cyclohexane formation. Molecular modeling studies show that (1 → 6) cyclization took place due to adopting a different catalytically competent docking pose of FPP derivatives compared to natural FPP within the active site of the BcBOT2, which is mainly due to the (2,3-Z)-configuration. This docking pose leads to a C1-C6 distance shorter (3.3-3.5 Å) than the typical association with a near-attack conformation required for cyclization. Finally, these biotransformation results were benchmarked by a comprehensive study of the “hydrolysis” of eight different FPP derivatives. Under enzyme-free conditions, cyclohexene and cycloheptene terpenoids are formed. The ring size is mainly determined by the position of the methyl group at C6 or C7. The study reveals that in addition to protein engineering, unnatural substrates can also be used to specifically manipulate the mode of cyclization of terpene synthases.
AB - The fungal sesquiterpene synthase BcBOT2 shows unique substrate promiscuity. It transforms farnesyl pyrophosphate (FPP) into presilphiperfolan-8β-ol via a cationic cascade that is initiated by a (1 → 11) cyclization. Here, it is shown that BcBOT2 also accepts (2,3-Z)-configured FPP derivatives, which provide terpenoids that result from an initial (1 → 6) cyclization. “Methyl mapping” was conducted by shifting the position of one or more methyl groups, and it was found that the location of methyl groups has a profound effect on the efficacy of cyclizations. In particular, the shift of the methyl group at C3 to the C2 position has the most profound effect on cyclohexane formation. Molecular modeling studies show that (1 → 6) cyclization took place due to adopting a different catalytically competent docking pose of FPP derivatives compared to natural FPP within the active site of the BcBOT2, which is mainly due to the (2,3-Z)-configuration. This docking pose leads to a C1-C6 distance shorter (3.3-3.5 Å) than the typical association with a near-attack conformation required for cyclization. Finally, these biotransformation results were benchmarked by a comprehensive study of the “hydrolysis” of eight different FPP derivatives. Under enzyme-free conditions, cyclohexene and cycloheptene terpenoids are formed. The ring size is mainly determined by the position of the methyl group at C6 or C7. The study reveals that in addition to protein engineering, unnatural substrates can also be used to specifically manipulate the mode of cyclization of terpene synthases.
KW - biotransformations
KW - computational modeling
KW - sesquiterpene
KW - sesquiterpene synthases
KW - terpenoids
UR - http://www.scopus.com/inward/record.url?scp=105004037930&partnerID=8YFLogxK
U2 - 10.1021/acscatal.5c01224
DO - 10.1021/acscatal.5c01224
M3 - Article
AN - SCOPUS:105004037930
VL - 15
SP - 8125
EP - 8139
JO - ACS catalysis
JF - ACS catalysis
SN - 2155-5435
IS - 10
ER -