The Homoserine Dehydrogenase thrA Significantly Impairs Biofilm Formation, Stress Adaptation, and Virulence in Klebsiella pneumoniae

Abstract

Klebsiella pneumoniae (K. pneumoniae) is a major opportunistic pathogen associated with a broad spectrum of hospital-and community-acquired infections. Biofilm formation contributes to bacterial persistence, stress tolerance, and host immune clearance. In this study, we constructed a transposon mutant library of the clinical K. pneumoniae strain KP20 using a mariner-based mutagenesis system and screened for mutants exhibiting defective biofilm formation. Targeted knockout of thrA significantly reduced biofilm formation and attenuated the adhesion and invasion of K. pneumoniae with respect to human airway epithelial Calu-3 cells and lung adenocarcinoma A549 cells. These phenotypic changes are associated with altered transcription of the gene clusters responsible for type I and type III fimbria biosynthesis. Moreover, deletion of thrA compromised bacterial tolerance to acidic and alkaline stresses and increased susceptibility to phagocytosis by dendritic cells. In a murine infection model, thrA deletion significantly attenuated the virulence of K. pneumoniae and decreased bacterial colonization in target organs. Collectively, these findings indicate that thrA substantially contributes to biofilm formation, pH stress tolerance, host–cell interaction, and virulence in K. pneumoniae.