The ease to mineralize organophosphates to release inorganic phosphorus (Pi) for plant uptake largely depends on the stereochemical structure of the compound, the types and sources of enzymes involved in addition to certain environmental factors. We determined the kinetics parameters (V_max, K_m, K_cat, and K_cat/K_m) of phosphomonoesterase and phytase from various sources on seven substrates analog  [p-Nitrophenyl phosphate, Bis-P-Nitrophenyl phosphate, P-Nitrophenyl phosphate bis(cyclohexylammonium), P-Nitrophenyl phosphate di(2-amino-2-ethyl-1-3-propanediol), D-Glucose 6-phosphate sodium salt, D-Glucose 6-phosphate disodium salt, and inositol hexakisphosphate] to assess the hydrolytic respond under standard conditions. Results from this study revealed that the V_max, K_m (the binding affinity), K_cat (turnover number) and K_cat/K_m (catalytic efficiency) values were different for similar enzyme from different sources. Similar trend was observed for E_a and Q₁₀. This study suggests that the optimal condition for enzyme activity is highly dependable on the substrates. The study enhances our understanding of the biogeochemical cycling of organophosphates in the environment.