Fig. 6: Characterization of LctB_A, LctE and LctC.

a Relative activities of the A domain in LctB with Leu or OH-Leu in the presence of LctE (left) and with Leu in the absence of LctE (right). Error bars indicate standard deviation (SD) (n = 3), and the data are presented as mean values ± SD. Black dots denote individual data points. b HPLC analysis (detected with absorption at 210 nm) of PCP domains in reaction mixtures (i) containing full components (LctB_A domain, LctE and Leu), (ii) without the LctB_A domain, (iii) without LctE, and (iv) with OH-Leu. A mass of 14533.71 corresponding to Leu-PCP (calculated mass, 14533.19) was detected (Figure S12). c The chemical structure of 4’ (left) and LC-ESI-MS (right, positive ion mode monitored at m/z 198.1) analysis of the reaction mixture using 4’ as the substrate (i) with LctC and (ii) without LctC. A peak with an m/z of 198.1130 (m/z, [M + H]⁺) corresponding to the exact mass of 6 (198.1125; m/z, [M + H]⁺) was observed in (i) (Fig. S13).