Aeromonas veronii is an important pathogen found in various aquatic environments and products, posing a threat to public health. The Hanks‐like serine/threonine protein kinase is closely linked to the pathogenesis of pathogenic bacteria, but the exact role of YihE in A. veronii remains still unknown. To study the specific function of the YihE kinase, we constructed a knockout mutant of the yihE gene in A. veronii. The deletion of the yihE gene resulted in changes to the metabolism of L‐arginine‐AMC and acetic acid, as well as enhanced resistance to ampicillin and kanamycin in A. veronii. Additionally, the ΔyihE strain demonstrated a 1.4‐fold increase in biofilm formation ability and a 1.8‐fold decrease in adhesion and invasion to EPCs when compared to the wild‐type strain. A significant decrease in cytotoxicity was observed at 2 and 3 h post‐infection with EPCs compared to the wild‐type strain. Additionally, the deletion of the yihE gene was associated with a significant decrease in motility of the strain. Furthermore, the deletion of the yihE gene resulted in a 1.44‐fold increase in the LD50 of A. veronii in zebrafish. These findings offer valuable insights into the pathogenic mechanisms of A. veronii.