Abstract:
Despite advancements in drug manufacturing and production, the purity of manufactured drugs and/or chemical compounds remains a critical challenge that requires further investigation. Enzyme engineering has emerged as a powerful tool for enhancing biocatalytic efficiency, stability, and substrate specificity, addressing key challenges in industrial and pharmaceutical applications. In this study, we tried to engineer AaDH to improve its catalytic performance in the amination reaction of industrially important chiral precursors (chemical compounds that are used as a building block for complex drugs) at a specific position in the chemical structure, such that the enzyme gives a specific enantiomer (enantioselective) as the product with very high purity that can be used in the industry. We used computational techniques like structural modelling and advanced techniques like Molecular Docking and Molecular Dynamics to study the enzyme’s active site and its probability of accepting our substrate of interest. The structural analysis using these computational techniques gave us critical insights into the active site of our wild-type enzyme and the mutations in the wild-type enzyme that are necessary to change the substrate scope of the enzyme. Further structural revealed that there is a list of residues that, when mutated, can increase the catalytic activity of the enzyme. Finally, we tried to clone and express the enzyme in Escherichia coli so that it could be tested and optimized further. We have proposed a final set of mutations that are necessary to increase the catalytic activity of AmDH.
