Figure 5
Enzymatic conversion of bafilomycin A1 to C1 using BfmI and BfmJ. (a) HPLC–MS chromatograms of ethyl acetate extracts of the enzymatic reactions. HPLC conditions were as follows: column: CAPCELL PAK C18, IF (2 μm, 2.0φ × 50 mm); mobile phase: 80% v/v methanol in 0.1% v/v formic acid; flow rate: 0.4 ml min−1. Extracted ion current (XIC) chromatograms are represented at m/z 645.397±0.013 (blue line) corresponding to bafilomycin A1 (m/z 645.397 [M+Na]+) and 743.397±0.013 (red line) corresponding to bafilomycin C1 (m/z 743.397 [M+Na]+), respectively. Each component was verified by authentic samples of bafilomycins A1 and C1. (b) 12.5% SDS-polyacrylamide gel electrophoresis of purified N-His8-tagged BfmI (left), N-His8-tagged BfmJ (right) and co-purified N-His6-tagged BfmI and intact BfmJ (center). A full color version of this figure is available at The Journal of Antibiotics journal online.
