Author(s) :

Aarnav S. Padigala.

Volume/Issue :

Abstract :

The growing issue of antibiotic resistance is causing an increasing crisis in bacterial infections. The discovery of new antibiotics is not aligned with the rapid increase in antibiotic resistance, resulting in significant challenges in treating antibiotic-resistant bacterial infections. One important bacterial pathogen with increased antibiotic resistance is Haemophilus influenzae, which was highlighted in the 2024 WHO bacterial priority pathogen list, especially in terms of the rise in resistance against ampicillin and other penicillin-based antibiotics. This study aimed to computationally characterize and compare the sequence, structure, and cellular localization of TEM-1 and ROB-1 β-lactamases in H. influenzae. In this study, multiple computational tools, including BLAST, Interpro, Phobius, Protein Data Bank, AlphaFold, and PyMOL, were used to investigate the prevalence of the most important genes and proteins associated with increased resistance to penicillin-based antibiotics in Haemophilus influenzae, specifically TEM-1 and ROB-1. BLASTn and BLASTp revealed a range of bacterial species, including those with intrinsic resistance, acquired resistance, and species with no documented ampicillin resistance, which were identified in the analysis as containing both genes and their associated proteins, thereby confirming the involvement of TEM-1 and ROB-1 in ampicillin resistance, as well as the widespread dissemination of TEM-1 and ROB-1-mediated resistance. A structural comparison of the two proteins revealed high similarity, and an Interpro analysis of protein domains revealed two distinct domains in both ROB-1 and TEM-1. Phobius analysis of protein cellular localization revealed that the two proteins are noncytoplasmic. These findings were in line with the mechanism of action of ampicillin resistance.