Identification of oligopeptide permease (opp) gene cluster in Vibrio fluvialis and characterization of biofilm production by oppA knockout mutation.
Oligopeptides play important roles in bacterial nutrition and signaling. The oligopeptide permease (opp) gene cluster was cloned from Vibrio fluvialis. The V. fluvialis opp operon encodes five proteins: OppA, B, C, D and F. The deduced amino acid sequence of these proteins showed high similarity with those from other Gram-negative bacteria. To investigate whether OppA is involved in biofilm production, an oppA knockout mutant was constructed by homologous recombination. The oppA mutant produced more abundant biofilm than the wild type in BHI medium. When both strains were grown in minimal medium, we could not detect biofilm formation. However, it was found that the biofilm productivity of the oppA mutant was two folds greater than that of the wild type in minimal medium containing peptone or tryptone. This variation in biofilm production was demonstrated by scanning electron microscopy (SEM). In minimal medium containing C-sources, both strains produced some biofilm without significant difference in the biofilm productivity. Complementation of oppA gene with the plasmid pOAC2, which contains oppA ORF plus promoter regions, was sufficient to restore growth rate and biofilm to the wild type. These results suggest that the OppA protein is involved in uptake of peptides and affects biofilm productivity.
Lee
Eun-Mi
EM
Department of Biotechnology and Bioengineering, Pukyong National University, Busan 608-737, Republic of Korea.
Ahn
Sun-Hee
SH
Park
Je-Hyun
JH
Lee
Jong-Hee
JH
Ahn
Soon-Cheol
SC
Kong
In-Soo
IS
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D018441:Biofilms; D003001:Cloning, Molecular; D026901:Membrane Transport Proteins / Q000235:genetics*; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D005810:Multigene Family; D016296:Mutagenesis; D014733:Vibrio / Q000201:enzymology* / Q000235:genetics* / Q000648:ultrastructure
D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; C037230:oligopeptide permease, Bacteria