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Localisation of alkaline phosphatase in the pore structure of paper

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Abstract

We study the bulk pore structure of bioactive paper impregnated with an enzyme, alkaline phosphatase (ALP), retained with a cationic polyacrylamide (CPAM) with synchrotron small-angle x-ray scattering and examine the translational dynamics of water in the porous material using 1H pulsed field gradient NMR. The aim of this study is to understand the perturbation of enzyme kinetics from the bulk behavior through the localization of the enzyme within the pore structure. We interpret the small-angle x-ray scattering data where the range of length scales examined in the scattering experiment corresponds to the packing of the unitary crystallites, or microfibrils, and aggregates of these cellulosic microfibrils. Pulsed field gradient NMR measurement of H2O diffusion in the pores between these aggregates indicates that some of these pores are strongly connected allowing convective mass transport, while others exhibit strongly restricted diffusion. Diffusion is also strongly anisotropic. We conclude that the CPAM retention polymer and the protein coat interior surfaces inside the pores of a particular size and that there is no major modification of the pore structure of paper by the CPAM or the ALP. The transport properties of this porous matrix are discussed in terms of the mechanisms by which sorption of ALP may perturb enzyme kinetics.

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Acknowledgements

Many thanks to Dr. Nathan Cowieson for expertise on the Synchrotron and to the Australian Synchrotron for beam time. CJG would like to thank Dr. Bogdan Chapman and Professor Philip Kuchel of the University of Sydney’s School of Molecular Biosciences for assistance with NMR measurements.

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Authors and Affiliations

  1. Australian Nuclear Science and Technology Organization (ANSTO), Locked Bag 2001, Kirrawee DC, NSW, 2232, Australia

    Christopher J. Garvey

  2. BioPRIA, Department of Chemical Engineering, Monash University, Clayton, VIC, 3800, Australia

    Mohidus Samad Khan & Gil Garnier

  3. Department of Chemical Engineering, Bangladesh University of Engineering and Technology, Dhaka, Bangladesh

    Mohidus Samad Khan

  4. Department of Physics and Astronomy, The University of Sheffield, Hicks Building, Hounsfield Road, Sheffield, S3 7RH, UK

    Michael P. Weir

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  1. Christopher J. Garvey
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Correspondence to Christopher J. Garvey.

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Garvey, C.J., Khan, M.S., Weir, M.P. et al. Localisation of alkaline phosphatase in the pore structure of paper. Colloid Polym Sci 295, 1293–1304 (2017). https://doi.org/10.1007/s00396-017-4037-5

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