Fig. 1
Mutant screening process. A Experimental procedure for screening mutants by modelling and simulating protein sequences and validating them in experiments. B RMSD of proteins at different ethanol concentrations and different pressures (10%, 50%, 100% ethanol concentrations, 1, 500, 1000 bar). C Compression coefficients of individual protein structures at different ethanol concentrations and different pressures. Each group represented a different loop structure, each line in the group represented a different pressure in the simulated environment, and each point on the line represented βT at different ethanol concentrations. D Heat map of free energy changes at mutant sites obtained from multi-scale free energy calculations, the red box in the figure is the final selected results of the research