Tweaking Enzymatic Electric Field Can Evolve CYP450-like Activity in Catalase and CYP-P411 Enzymes.

Unlike CYP450 (cysteine-ligated), the axial ligations in Catalase (tyrosine-ligated) and CYP411-BM3 (serine-ligated) are different, which is generally attributed to the different catalytic efficiencies of these enzymes. Studies show that an interplay between push-pull effects due to different ligations modulates the reactivity of different enzymes. In the present study, we employ extensive MD simulation and a hybrid QM/MM approach to demonstrate that differences in the axial ligation in conjunction with protein residues generate a preorganized local electric field (LEF) that, in turn, controls the reactivity of the different families of heme-containing enzymes. Considering an example of the epoxidation reaction in trans-β-methylstyrene, we show that, by strategically mutating catalase and CYP411-BM3 to design a preorganized LEF that matches the native LEF of CYP450 monooxygenase, we can evolve catalase and CYP411-BM3 with reactivity similar to that of CYP450. This study shows that by tuning the preorganized LEF of an enzyme through strategic mutations of charged and polar residues, a secondary, non-native function can be evolved.