Because of our limited understanding of the mechanisms governing Vibrio species persistence and spread in the face of global warming, finding ways to control the increasing spread of pathogenic vibrios is challenging. To determine whether the persistence of Vibrio harveyi is associated with physiological and gene expression patterns, it is important to do research on several proteins in its genome which are classified as hypothetical proteins (HPs). As a result, the current work sought to elucidate the roles of a HP found in the genome of V.harveyi. To determine the structure, function and homologous model of this protein, quality bioinformatics methods were used to predict and confirm the function as well as secondary and tertiary structure. Additionally, the active site and interacting proteins were examined using CASTp and the STRING server. An important biological activity of the HP is that it contains single functional domains that may be act as DNA binding site. Further, protein-protein interactions within selected HP revealed several functional partners that are essential for bacterial survival with different functional activity. In addition, molecular docking and simulation results showed stable bonding between HP and ExbD/TolR family protein which might be of significant relevance to future bacterial genetics research.