Evaluation of biochemical and molecular polymorphism in extended spectrum β-lactamases of Mycobacterium tuberculosis clinical isolates

Background Tuberculosis (TB) caused 1.8 million deaths worldwide with increased multiple drug resistance (MDR) cases estimated 4.8 lakhs in the year 2015. β-Lactam antibiotics could be a hope for TB treatment. Therefore, in this study, uniformity in the biochemical and molecular nature of β-lactamases was analyzed to evaluate the potential of β-lactam antibiotics as a treatment regimen against Mycobacterium tuberculosis (MTB). Materials and methods β-Lactamase enzymes in 233 MTB clinical isolates along with control H37Rv strain were characterized by enzyme kinetic using nitrocefin and cefotaxime as a substrate, isoelectric points by isoelectric focusing electrophoresis (IEF) and by PCR and Southern blotting. Results Enzyme kinetics showed K m and V max for nitrocefin in the range of 56-69μM and 7.00-11IU/lit respectively, for cefotaxime in the range of 0.35-0.59μM and 18-25IU/lit respectively. β-Lactamase showed high affinity for clavulanic acid an inhibitor of Extended-Spectrum β-lactamase enzymes (ESBLs). The pIs of 4.9 and 5.1 were observed for all the MTB clinical isolates and control H37Rv. Southern blotting confirmed the presence of blaC sequence in MTB chromosomal DNA. Conclusion This confirmed that MTB β-lactamase enzymes belong to the Class A, group 2be Extended Spectrum β-Lactamases with no biochemical or molecular polymorphism. ESBLs are mainly responsible for resistance against β-lactam antibiotics in MTB. Thus ESBLs could be the potential therapeutic target for TB treatment using β-lactam antibiotics in combination with β-lactamase inhibitors like sulbactam and sodium clavulanate.