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- Title
Molecular insights into the catalytic promiscuity of a bacterial diterpene synthase.
- Authors
Li, Zhong; Zhang, Lilan; Xu, Kangwei; Jiang, Yuanyuan; Du, Jieke; Zhang, Xingwang; Meng, Ling-Hong; Wu, Qile; Du, Lei; Li, Xiaoju; Hu, Yuechan; Xie, Zhenzhen; Jiang, Xukai; Tang, Ya-Jie; Wu, Ruibo; Guo, Rey-Ting; Li, Shengying
- Abstract
Diterpene synthase VenA is responsible for assembling venezuelaene A with a unique 5-5-6-7 tetracyclic skeleton from geranylgeranyl pyrophosphate. VenA also demonstrates substrate promiscuity by accepting geranyl pyrophosphate and farnesyl pyrophosphate as alternative substrates. Herein, we report the crystal structures of VenA in both apo form and holo form in complex with a trinuclear magnesium cluster and pyrophosphate group. Functional and structural investigations on the atypical 115DSFVSD120 motif of VenA, versus the canonical Asp-rich motif of DDXX(X)D/E, reveal that the absent second Asp of canonical motif is functionally replaced by Ser116 and Gln83, together with bioinformatics analysis identifying a hidden subclass of type I microbial terpene synthases. Further structural analysis, multiscale computational simulations, and structure-directed mutagenesis provide significant mechanistic insights into the substrate selectivity and catalytic promiscuity of VenA. Finally, VenA is semi-rationally engineered into a sesterterpene synthase to recognize the larger substrate geranylfarnesyl pyrophosphate. Diterpene synthase VenA catalyses the synthesis of venezuelaene A with a unique 5-5-6-7 tetracyclic skeleton from geranylgeranyl pyrophosphate. Here, the authors report crystal structures of apo- and holo-VenA, provide mechanistic insights into its substrate selectivity and promiscuity, and engineer VenA into a sesterterpene synthase.
- Subjects
TERPENES; ENGINEERS; CRYSTAL structure; DITERPENES; SYNTHASES; MAGNESIUM compounds; MUTAGENESIS
- Publication
Nature Communications, 2023, Vol 14, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-023-39706-9