Fig. 5: Structural investigations of MicA.

a Investigations utilized a tFold-based model of MicA. The structural model of MicA highlights key active site residues within a 5 Å distance, and the docked GGPP, depicted in stick, exhibits a pre-cyclization conformation. b The docking results facilitated the measurement of distances from C1 to C10 and C1 to C14. The distances are shown in Å. c Comparative analysis of relative activities of wild-type MicA (red) and its mutants (blue). The yield of 1 was quantified, establishing the relative activity of wild-type MicA at 100%. Three biological parallel (n = 3) replicates were performed for both wild-type MicA and its mutants, with the relative activities of each replicate indicated by triangular points. The data are presented as mean values plus the standard error of the mean (SEM). Compared to MicA, all relative activities of mutants have p values less than 0.0001, except for MicA^(E53W) with a value of 0.0025. Source data are provided as a Source Data file. The “♯”shown denotes mutants that can produce new compounds. d HPLC traces (210 nm) of activities of wild-type MicA, MicA^(V220A), and MicA^(L221A), The term “EV” refers to an “empty vector” and products marked with asterisks (*) were uncharacterized in this study. e The chemical structures of the diterpenes synthesized by MicA^(V220A) and MicA^(L221A) are presented.