TerDockin data for activity prediction and engineering of mutants of Bradyrhizobium japonicum ent-kaurene synthase (BjKS)

Provided here are the score files output by the TerDockin Rosetta protocol when generating docked structures of BjKS with various intermediates related to the production of ent-kaurene. In order to asses the docking protocol when considering the targeting of production of ent-pimara-8,15-diene, a previously identified mutant that produces this compound was tested. Here a comparison of two possible intermediates that could yield this product are tested (C8 vs C9) before using the winning intermediate to retrospectively examine the activity of BjKS F72Y. To further improve specificity of BjKS for this product, an in silico serine threonine scan of the active site was conducted using TerDockin to identify other sites likely to produce ent-pimara-8,15-diene. Building on identified producers of the targetted compound, a further mutant set comprising G32X Y280X F72Y I166F is explored. The results from this iteration yielded a final pair of mutation sites, F72X Y280X, for TerDockin analysis. Overall, these data enabled the discovery of 10 mutants of BjKS which show specificity for ent-pimara-8,15-diene with F72Y Y280S showing over 90% specificity for this product.