Presentation of functional organophosphorus hydrolase fusions on the surface of Escherichia coli by the AIDA-I autotransporter pathway.
We report, the surface presentation of organophosphorus hydrolase (OPH) and green fluorescent protein (GFP) fusions by employing the adhesin-involved-in-diffuse-adherence (AIDA-I) translocator domain as a transporter and anchoring motif. The surface location of the OPH-GFP fusion protein was confirmed by immunofluorescence microscopy, and protease accessibility, followed by Western blotting analysis. The investigation of growth kinetics and stability of resting cultures showed that the presence of the AIDA-I translocator domain in the outer membrane neither inhibits cell growth nor affects cell viability. Furthermore, the surface-exposed OPH-GFP was shown to have enzymatic activity and a functional fluorescence moiety. These results suggest that AIDA-I autotransporter is a useful tool to present heterologous macromolecule passenger proteins on the bacterial surface. Our strategy of linking GFP to OPH and the possibility to employ various bacterial species as host has enormous potential for enhancing field use.
Li
Chaokun
C
State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China.
Zhu
Yaran
Y
Benz
Inga
I
Schmidt
M Alexander
MA
Chen
Wilfred
W
Mulchandani
Ashok
A
Qiao
Chuanling
C
D018830:Adhesins, Escherichia coli / Q000502:physiology*; D043303:Aryldialkylphosphatase / Q000235:genetics / Q000378:metabolism*; D001673:Biodegradation, Environmental; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000737:chemistry; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D010755:Organophosphates / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*
C075559:AIDA-I protein, E coli; D018830:Adhesins, Escherichia coli; D008565:Membrane Proteins; D010755:Organophosphates; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D043303:Aryldialkylphosphatase