In Silico Functional Annotation of Hypothetical Proteins from Burkholderia pseudomallei strain GTC3P0254T

Burkholderia pseudomallei , the causative agent of melioidosis, contains numerous hypothetical proteins (HPs) with unknown functions, limiting our understanding of its biology and pathogenicity. This study employed in silico approaches to functionally annotate 27 HPs from B. pseudomallei strain GTC3P0254T using domain analysis, physicochemical characterization, structural modeling, and protein-protein interaction predictions. The identified HPs were classified into enzymes, transporters, binding proteins, regulatory proteins, and structural proteins. Notably, several HPs exhibited enzymatic activity, including polyphosphate kinase and isoprenoid synthase, which play crucial roles in bacterial metabolism and survival. Additionally, membrane-associated proteins were linked to drug resistance and host adaptation, while one HP demonstrated ubiquitin hydrolase activity, a function associated with bacterial invasion and virulence. Homology-based tertiary structure predictions were validated using multiple structural assessment tools, and protein-protein interaction analyses provided insights into their functional associations. These findings enhance our understanding of B. pseudomallei pathogenesis and antimicrobial resistance, highlighting potential targets for therapeutic interventions. However, since this study is based solely on computational predictions, experimental validation through biochemical assays and genetic studies is essential to confirm these findings. Future research should explore these HPs as potential drug targets and diagnostic biomarkers to improve treatment strategies for melioidosis.