Following from this, the LFER shown in Figure provides evidence of the maturity of computational approaches and their power to operate alongside experimental studies in moving towards a comprehensive explanation of the catalytic rate acceleration for TIM and other enzymes. It has been proposed that the prime imperative for enzymatic catalysis of deprotonation of weak carbon acids is that protein catalysts operate on the bound substrates to reduce that standard-state Gibbs free energy for deprotonation of carbon. type TIM and all TIM variants described in this study. The correlation coefficients were calculated using regression analysis to be 0.994 and 0.995 for the deprotonation of substrates DHAP and GAP respectively. In this figure, the black circles correspond to the non-enzymatic reaction in aqueous solution, the blue circles correspond to the wild-type enzyme, the red circles correspond to single substitutions of polar side chains (K12G and E97A), the green circles correspond to single substitutions of nonpolar side chains (P166A, I170A and L230A), and finally, the orange circle corresponds to double substitutions of nonpolar side chains (I170A/L230A). The raw data used for this figure is provided in Table .