The impact of the flagellar protein gene fliK on Helicobacter pylori biofilm formation

The biofilm structure of Helicobacter pylori is known to enhance its capabilities for antimicrobial resistance. This study aims to investigate the role of the flagellar hook length control protein gene fliK in the biofilm formation of H. pylori. Homologous recombination was employed to knock out the fliK gene in the H. pylori NCTC 11637 strain. The flagella of H. pylori were observed using transmission electron microscopy (TEM), whereas H. pylori motility and growth were examined through semi-solid agar assays and growth curve analyses, respectively. The bacterial biofilm and its constituents were visualized utilizing fluorescence confocal microscopy. Assessments of H. pylori adhesion to gastric mucosal cells, its vacuolar toxicity, and antibiotic resistance were evaluated using co-culture experiments and E-test methods. The fliK gene was successfully knocked out in H. pylori NCTC 11637. The Delta fliK mutant exhibited polyhook structures or lacked typical flagellar morphology, reduced mobility, and a slower bacterial growth rate compared with the wild-type strain. Fluorescence confocal microscopy revealed a decrease in the thickness of the biofilm formed by the Delta fliK strain, along with reductions in polysaccharide and DNA components. The deletion of fliK did not affect vacuolar toxicity or antibiotic resistance but did reduce the adhesive capacity of the bacterium to gastric mucosal cells. The deletion of the fliK gene significantly impairs H. pylori biofilm formation, leading to substantial decreases in biofilm components, bacterial growth, and adhesion capabilities. These findings underscore the importance of fliK in the pathogenicity of H. pylori.