Methanol induced inactivation of Dieselzymes. (A) Methanol inactivation as a function of concentration. Samples were incubated for 2 hours and assayed after dilution to 10% methanol. Results are the average of 3 independent experiments. (B) Methanol inactivation as a function of time. Samples were incubated in 50% methanol for 24 hours. At indicated time points, samples were diluted to 10% methanol and assayed for residual activity. Error-bars are omitted but are less than 5% in all cases. Lines are shown for clarity only. Results are the average of 3 independent experiments. WT PML (black circles), BCL (red open circles), and Dieselzyme 1 (blue squares), Dieselzyme 2 (blue diamond), Dieselzyme 3 (green triangle), and Dieselzyme 4 (orange inverted triangle). (C) Inactivation profile of Wild-Type PML (black bar), Dieselzyme 4 (orange bar), and BCL (white bar) in response to various solvents. Enzymes were incubated for 24 hours in 70% solvent and diluted to 10% prior to assaying for residual activity. Results are the average of three independent experiments.