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Cited by 1Plant biochemistry and biosynthesisMicrobial Natural Products and BiosynthesisEnzyme Catalysis and Immobilization

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Sequence-Guided Engineering of a Bacterial Diterpene Synthase Induces Further Cyclization

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Wei, Xiuting, Ning, Wenbo, Huang, Pin-Shuo, Nafie, Jordan, Ghiviriga, Ion, Rudolf, Jeffrey D.·2026·ACS Catalysis
DOI 10.1021/acscatal.6c04391PMC PMC13557768

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Abstract

Enzyme engineering serves as a powerful tool in biocatalysis, enabling the development of enzymes with improved stability, activity, and specificity for a range of academic, industrial, and pharmaceutical applications. However, a limited understanding of sequence–structure–function relationships in terpene synthases, the enzymes that form the complex polycyclic hydrocarbon skeletons of terpenoid natural products, presents a major challenge in predicting and engineering the products of terpene synthases. In this study, we investigated the product profiles of two bacterial variediene synthases, OdVS from Olivibacter domesticus and PsVS from Prauserella shujinwangii, and found that they share some products with the bacterial phomopsene synthase, PmS from Allokutzneria albata, despite low sequence similarity. Thirteen diterpenes were isolated and structurally elucidated including two previously unreported compounds. A series of variants of OdVS, PsVS, and PmS were constructed by targeting conserved residues around the active site and aided in the identification of key residues that control the cyclization pathway. Ultimately, mutation of a single residue, PsVSY86L, was found to switch the major product of PsVS from the tricyclic variediene to the tetracyclic phomopsene, although this switch came at the cost of significantly reduced overall yield indicating a tradeoff between activity and product diversification.

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Metadata

Authors
Wei, X., Ning, W., Huang, P.-S., Nafie, J., Ghiviriga, I., & Rudolf, J. D.
Affiliations
  • University of Florida (US)
  • BioTools, Inc. , 5730 Corporate Way, Suite 220 , , ,
Corresponding author
Rudolf, Jeffrey D.
ORCID
Venue
ACS Catalysis
Publisher
American Chemical Society (ACS)
Published
2026-08-24
Volume
16
Issue
17
Pages
17275-17283
ISSN
2155-5435 (ISSN-L)
DOI
10.1021/acscatal.6c04391
PMC
PMC13557768
OpenAlex
W7163692377
Cited by
1
References
1
Citations by year
2026: 1
FWCI
2.07
Type
journal article
Language
en
Open access
Licence
Locations
  • ACS Catalysis (publishedVersion)
  • ChemRxiv (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • NIH Office of the Director (OD021758-01A1)
  • National Institute of General Medical Sciences (R35 GM142574)
  • University of Florida
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant biochemistry and biosynthesis
  • Life Sciences › Immunology and Microbiology › Applied Microbiology and Biotechnology › Microbial Natural Products and Biosynthesis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Enzyme Catalysis and Immobilization
Keywords
diterpene synthase, enzyme engineering, terpene cyclization, product profile, sequence-structure-function relationships, active site residues, residue substitution, tetracyclic diterpenes, biocatalysis

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CoChat. (2026). Citation check: Sequence-Guided Engineering of a Bacterial Diterpene Synthase Induces Further Cyclization. https://app.cochat.ai/papers/10.1021/acscatal.6c04391
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Wei, X., Ning, W., Huang, P.-S., Nafie, J., Ghiviriga, I., & Rudolf, J. D. (2026). Sequence-Guided Engineering of a Bacterial Diterpene Synthase Induces Further Cyclization. ACS Catalysis. https://doi.org/10.1021/acscatal.6c04391