Extended Data Fig. 2: DMSP cleavage activity of purified DddL.
From: Acrylate protects a marine bacterium from grazing by a ciliate predator
a, SDS-polyacrylamide gel electrophoresis assay of the purified DddL protein. The DddL protein is indicated by a black arrow. M, protein molecular weight marker. Each picture is a representative of at least three repeats. b, SDS-polyacrylamide gel electrophoresis and western blot assays of the overexpression profile of DddL in the membrane and the cytoplasmic fractions. Marker, protein molecular weight marker with the unit of kDa. Me, proteins in the membrane; Cyto, proteins in the cytoplasm; DddL, the purified DddL. WB, western blot. White arrows indicate the DddL protein band. Each picture is a representative of at least three repeats. c, Detection of the DMSP cleavage activity of the purified DddL protein by measuring the production of acrylate via HPLC. The black arrow indicates the absorption peak of acrylate at 214 nm. The dashed line presents the direction of z-axis. d, The effect of pH on DddL activity. The activity of DddL at pH 9.0 was defined as 100%. Data are presented as mean values +/- standard deviation. Error bars represent standard deviation of triplicate experiments. e, The effect of temperature on DddL activity. The activity of DddL at 30 °C was defined as 100%. Data are presented as mean values +/- standard deviation. Error bars represent standard deviation of triplicate experiments. f–h, Enzymatic kinetic parameters for DMSP cleavage by DddL. A linear fit curve measured at 4 °C, pH 8.0 (f), and non-linear fit curves measured at 18 °C, pH 8.0 (g), and 30 °C, pH 9.0 (h).
