— A pseudo-reverse approach is presented in this paper to analyze the evolutionary behaviour of enzymes. It employs the standard model of Nei and Gojobori [1] in a generalized form for determining the nucleotide substitutions and Jukes and Cantor’s [2] model for finding the out their rates. Comparative genomics is also embedded in this model to calculate the lineages among the species like human, mouse and rat for these enzyme proteins. It is predicted from this study that the mutation for the enzymes are comparatively slower than ordinary proteins and the time of divergence for these enzymes with human and mouse or rat is almost five times more, around 400 Million years. Hence, this paper describes the methodology and the findings in details.