Cellobiose-Specific Phosphotransferase System of Klebsiella pneumoniae and Its Importance in Biofilm Formation and Virulence
Resource
Infect. Immun., 80(7), 2464-2472
Journal
Infection and Immunity
Pages
2464-2472
Date Issued
2012
Date
2012
Author(s)
Abstract
Klebsiella pneumoniae is a Gram-negative bacillus belonging to the family Enterobacteriaceae. In the past 20 years, K. pneumoniae has become the predominant pathogen causing community-acquired pyogenic liver abscess (PLA). The formation of biofilm facilitates bacterial colonization and has been implicated in reduced susceptibility to the host immune response. To investigate genes related to biofilm formation in a PLA-associated K. pneumoniae strain, a transposon mutant library was screened by microtiter plate assay to identify isolates impaired for biofilm formation. One of the mutants was disrupted in celB, encoding the putative cellobiose-specific subunit TIC of enzyme II (EIIC) of a carbohydrate phosphotransferase system (PTS). This transmembrane protein is responsible for recognizing and binding specific sugars and transporting them across the cell membrane into the cytoplasm. Deletion and chromosomal complementation of celB confirmed, by microtiter plate and slide culture assays, that celB was indeed responsible for biofilm formation. Cellobiose-specific PTS activities of deletion mutants grown in LB broth and 0.005% cellobiose minimal medium were markedly lower than that of the wild-type strain grown under the same conditions, thereby confirming the involvement of celB in cellobiose transport. In 0.005% cellobiose minimal medium, the celB mutant showed a delay in growth compared to the wild-type strain. In a mouse model of intragastric infection, deletion of the celB gene increased the survival rate from 12.5% to 87.5%, which suggests that the celB deletion mutant also exhibited reduced virulence. Thus, the celB locus of K pneumoniae may contribute to biofilm formation and virulence through the metabolism of cellobiose.
SDGs
Other Subjects
bacterial enzyme; cellobiose; membrane protein; phosphotransferase; sellobiose specific phosphotransferase system; unclassified drug; animal experiment; animal model; animal tissue; article; bacterial biofilm formation; bacterial gene; bacterial growth; bacterial phenomena and functions; bacterial virulence; cell culture; cell transport; controlled study; enzyme activity; female; gene deletion; human; human tissue; Klebsiella pneumoniae; microtiter plate assay; mouse; nonhuman; nucleotide sequence; priority journal; protein binding; pyogenic liver abscess; Animals; Biofilms; Cellobiose; Disease Models, Animal; DNA Transposable Elements; Female; Gene Knockout Techniques; Humans; Klebsiella Infections; Klebsiella pneumoniae; Liver Abscess; Mice; Mice, Inbred BALB C; Mutagenesis, Insertional; Phosphoenolpyruvate Sugar Phosphotransferase System; Survival Analysis; Virulence; Virulence Factors
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