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Display of Organophosphorus Hydrolase on the Cyanobacterial Cell Surface Using Synechococcus Outer Membrane Protein A as an Anchoring Motif

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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.

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Acknowledgments

The authors thank Chutirat Assawathep and Thatri Iampornsin for initial technical assistance and E for L International Co., Ltd. for the use of confocal laser microscope. This work was supported by Mahidol University Research Grant and The Thailand Research Fund to Wipa Chungjatupornchai.

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Authors and Affiliations

  1. Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Nakornpathom, 73170, Thailand

    Wipa Chungjatupornchai, Attapon Kamlangdee & Sirirat Fa-aroonsawat

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  1. Wipa Chungjatupornchai
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  2. Attapon Kamlangdee
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  3. Sirirat Fa-aroonsawat
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Correspondence to Wipa Chungjatupornchai.

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Chungjatupornchai, W., Kamlangdee, A. & Fa-aroonsawat, S. Display of Organophosphorus Hydrolase on the Cyanobacterial Cell Surface Using Synechococcus Outer Membrane Protein A as an Anchoring Motif. Appl Biochem Biotechnol 164, 1048–1057 (2011). https://doi.org/10.1007/s12010-011-9193-3

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  • DOI: https://doi.org/10.1007/s12010-011-9193-3

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