*3.12. Determination of Catalytic Parameters*

Kinetic parameters were calculated by plotting the rate of p-NPP and bis-p-NPP splitting at concentrations from 5 to 15 mM in a buffer containing 25 mM Tris-HCl, pH 9.0, 2 mM CoCl2, 2 mM FeCl3. The reaction was carried out at 25 ◦C. The Michaelis constant Km, the maximum reaction rate Vmax, and the turnover number kcat were determined by plotting the Layuver–Burk graph using the OriginPro 8.5 program.

#### *3.13. Molecular Modeling*

The target template alignment customization of the modeling process and 3D model building for CamPhoD (GenBank: WP\_043333989) were carried out using the Molecular Operating Environment (MOE) version 2018.01 package, using the Amber12: EHT forcefield (EHT-Extended Hueckel Theory) [62]. The alkaline phosphatase D from *B. subtilis* (PDB code: 2YEQ) was used as a template, which had a high-resolution crystal structure. The evaluation of structural parameters, contact structure analysis, physicochemical properties, and visualization of the results were carried out with the ligand interaction and dock modules in the MOE 2018.01 program [62].
