Using mechanism similarity to understand enzyme evolution.

Enzyme reactions take place in the active site through a series of catalytic steps, which are collectively termed the enzyme mechanism. The catalytic step is thereby the individual unit to consider for the purposes of building new enzyme mechanisms - i.e. through the mix and match of individual catalytic steps, new enzyme mechanisms and reactions can be conceived. In the case of natural evolution, it has been shown that new enzyme functions have emerged through the tweaking of existing mechanisms by the addition, removal, or modification of some catalytic steps, while maintaining other steps of the mechanism intact. Recently, we have extracted and codified the information on the catalytic steps of hundreds of enzymes in a machine-readable way, with the aim of automating this kind of evolutionary analysis. In this paper, we illustrate how these data, which we called the "rules of enzyme catalysis", can be used to identify similar catalytic steps across enzymes that differ in their overall function and/or structural folds. A discussion on a set of three enzymes that share part of their mechanism is used as an exemplar to illustrate how this approach can reveal divergent and convergent evolution of enzymes at the mechanistic level. Supplementary Information: The online version contains supplementary material available at 10.1007/s12551-022-01022-9.