Computational Evaluation of Designed Phosphatase from Conserved Sequence Scratch for Diverse Substrate Specificity

Abstract

The ability to design efficient enzymes for a broad class of different reactions would be of tremendous practical interest in both science and industry. Computer-assisted designing is a novel approach to generating industrial enzymes for biotechnological applications. Objectives: The main aim of this study was to design an enzyme construct with diverse substrate-binding specificity based on the evolutionary conservation of archaeal vanadium-dependent phosphatases. Materials and methods: A rational 3D structural model of enzyme construct was developed from conserved sequence scratch encompassing a vanadium-binding site and functional domain. Substrate-binding specificity of the designed enzyme was computed with different myo-inositol polyphosphate analogous by a molecular docking program. Results: A designed enzyme has shown more substrate-binding specificity with 1D-myo-inositol 3, 4, 5, 6-tetrakisphosphate. Its catalytic function closely resembled myo-inositol polyphosphate-5-phosphatase and multiple inositol polyphosphate phosphatases. Moreover, the enzyme construct was energetically stable with a low degree of conformational changes upon substrate-binding. Conclusion: Substrate specificity and catalytic competence of designed enzymes were computationally evaluated for further biotechnological applications.