Structure-Guided Product Determination of Bacterial Type II Diterpene Synthases

28 June 2022, Version 1
This content is a preprint and has not undergone peer review at the time of posting.


A grand challenge in terpene synthase (TS) enzymology is the ability to predict function from protein sequence. Given the limited number of characterized bacterial TSs and significant sequence diversities between them and their eukaryotic counterparts, this is currently impossible. To define the sequence-structure-function relationships of type II bacterial TSs, we determined the structure of the terpentedienyl diphosphate synthase Tpn2 from Kitasatospora sp. CB02891 by X-ray crystallography and made structure-guided mutants to probe its mechanism. Substitution of a glycine into a basic residue changed the product preference from the clerodane skeleton to a syn-labdane skeleton, resulting in the first syn-labdane identified from a bacterial TS. Understanding how a single residue can dictate cyclization patterns in bacterial type II TSs, along with detailed bioinformatics analysis of these TSs, sets the stage for the investigation of the functional scope of bacterial type II TSs and the discovery of novel bacterial terpenoids.


Terpene synthase
X-ray crystallography
Natural Products

Supplementary materials

Supporting Information for Tpn2
Supporting Data, Methods, and References.


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