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KMID : 0545120110210121330
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Journal of Microbiology and Biotechnology
2011 Volume.21 No. 12 p.1330 ~ p.1335
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Suppressing Erwinia carotovora Pathogenicity by Projecting N-Acyl Homoserine Lactonase onto the Surface of Pseudomonas putida Cells
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Qianqian Li
Hong Ni
Shan Meng
Yan He
Ziniu Yu
Lin Li
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Abstract
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N-Acyl homoserine lactones (AHLs) serve as the vital quorum-sensing signals that regulate the virulence of the pathogenic bacterium Erwinia carotovora. In the present study, an approach to efficiently restrain the pathogenicity of E. carotovora-induced soft rot disease is described. Bacillus thuringiensis-derived N-acyl homoserine lactonase (AiiA) was projected onto the surface of Pseudomonas putida cells, and inoculation with both strains was challenged. The previously identified N-terminal moiety of the ice nucleation protein, InaQ-N, was applied as the anchoring motif. A surface display cassette with inaQ-N/ aiiA was constructed and expressed under the control of a constitutive promoter in P. putida AB92019. Surface localization of the fusion protein was confirmed by Western blot analysis, flow cytometry, and immunofluorescence microscopy. The antagonistic activity of P. putida MB116 expressing InaQ-N/AiiA toward E. carotovora ATCC25270 was evaluated by challenge inoculation in potato slices at different ratios. The results revealed a remarkable suppressing effect on E. carotovora infection. The active component was further analyzed using different cell fractions, and the cell surface-projected fusion protein was found to correspond to the suppressing effect.
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KEYWORD
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Pseudomonas putida, Cell surface display, N-acyl homoserine lactonase, Erwinia carotovora, Antibacterial activity
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