Molecular and computational insights into algD biofilm genes in multi drug resistant and extensively drug resistant Pseudomonasaeruginosa.

Antibiotic-resistance and biofilm formation are the main virulence factors and present a serious treatment challenge in Pseudomonas aeruginosa. This study aimed to investigate antimicrobial resistance, genetic diversity, biofilm-specific algD gene, and computational analysis of clinical isolates. Forty two isolates of P. aeruginosa were examined by PCR, ELISA, sangers sequencing, phylogenetic analysis, MolProbity score, 3D structural modelling, Ramachandran plot, multiple sequence alignment, and protein domain analysis. According to the results, PCR analysis revealed algD gene presence in all isolates. ELISA showed 55 % (n = 23) of the samples produced strong biofilms, 38 % (n = 16) produced moderate biofilms, and 7 % (3) produced weak biofilms. The evolutionary relationships of 8 (S1-S8) P. aeruginosa strains with 81 reference strains were illustrated by the phylogenetic tree. Samples S1-S8 showed excellent MolProbity score (<1.00), low clashed scores (0.67-0.70), most residues in the favored regions (∼96.2-96.5 %), low Ramachandran outliers (0.53-0.56 %), low Rotamer outlier (0.62 %), low bad angles (<2), indicated high-quality models and values preferred percentages showed excellent models with structural refinement. Over all samples S5 and S6 stood out as the top choices for high-confidence modeling and applications. The essential catalytic domain UDP-glucose/GDP-mannose dehydrogenase was identified that could be used as important therapeutic targets. High quality models indicated suitability for downstream applications, such as studying protein-ligand interactions, understanding structural aspects of biofilm-resistant bacteria. This study improved our knowledge of the mechanisms underlying P. aeruginosa biofilm resistance and sets the stage for the development of novel therapeutic and diagnostic strategies to combat multidrug resistant strains.