Understanding and characterisation of BlaA beta-lactamase from Yersinia enterocolitica and in-silico characterization of MAB beta-lactamase from Mycobacterium abscessus

Abstract

The accidental discovery of antibiotic penicillin by Sir Alexander Fleming was termed a significant breakthrough in the history of humanity. What started as a revolutionary approach to treating diseases is now a cause of concern. Antimicrobial resistance is predicted to become one of the leading causes of death worldwide by 2050. One of the fundamental mechanisms responsible for antimicrobial resistance is the production of beta-lactamases. Therefore, understanding these enzymes and characterising them becomes of utmost importance for developing new drugs against them. This project involves in-vitro as well as in-silico characterisation of two of these beta-lactamases: BlaA from Yersinia enterocolitica and MAB from Mycobacterium abscessus. BlaA was purified, followed by biochemical and biophysical characterisation using Steady-state kinetics, Inhibition kinetics, Fluorescence spectroscopy, Circular Dichroism Spectroscopy, and Structure determination through X-ray diffraction. We used in-silico techniques for Sequence analysis, Structure-based analysis, Phylogenetic analysis, Molecular Docking, and Dynamic studies for MAB beta-lactamase.