title: Display of organophosphorus hydrolase on the cyanobacterial cell surface using synechococcus outer membrane protein a as an anchoring motif.
abstract: The display of proteins to cyanobacterial cell surface is made complex by combination of Gram-positive and Gram-negative features of cyanobacterial cell wall. Here, we showed that Synechococcus outer membrane protein A (SomA) can be used as an anchoring motif for the display of organophosphorus hydrolase (OPH) on cyanobacterial cell surface. The OPH, capable of degrading a wide range of organophosphate pesticides, was fused in frame to the carboxyl-terminus of different cell-surface exposed loops of SomA. Proteinase K accessibility assay and immunostaining visualized under confocal laser scanning microscopy demonstrated that a minor fraction of OPH with 12 histidines fused in frame with the third cell-surface exposed loop of SomA (SomAL3-OPH12H) was displayed onto the outermost cell surface with a substantial fraction buried in the cell wall, whereas OPH fused in frame with the fifth cell-surface exposed loop of SomA (SomAL5-OPH) was successfully translocated across the membrane and completely displayed onto the outermost surface of Synechococcus. The successful display of the functional heterologous protein on cell surface provides a useful model for variety of applications in cyanobacteria including screening of polypeptide libraries and whole-cell biocatalysts by immobilizing enzymes.
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Nakornpathom 73170, Thailand. stwcj@mahidol.ac.th
authors.forename: Wipa
authors.initials: W
authors.lastname: Chungjatupornchai
authors.forename: Attapon
authors.initials: A
authors.lastname: Kamlangdee
authors.forename: Sirirat
authors.initials: S
authors.lastname: Fa-Aroonsawat
mesh_terms: D020816:Amino Acid Motifs; D043303:Aryldialkylphosphatase / Q000235:genetics* / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D001673:Biodegradation, Environmental; D001709:Biotechnology / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism; D005417:Flavobacterium / Q000737:chemistry / Q000378:metabolism*; D007120:Immunochemistry; D007306:Insecticides / Q000378:metabolism*; D018613:Microscopy, Confocal; D009943:Organophosphorus Compounds / Q000378:metabolism*; D010957:Plasmids; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D046940:Synechococcus / Q000235:genetics* / Q000378:metabolism
chemical_list: D001425:Bacterial Outer Membrane Proteins; D007306:Insecticides; D009943:Organophosphorus Compounds; D011993:Recombinant Fusion Proteins; D043303:Aryldialkylphosphatase