Study into the Kinetic Properties and Surface Attachment of a Thermostable Adenylate Kinase

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0.1mM_ADP.xlsx

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0.05mM_ADP_PP.xlsx

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0.0016.xlsx

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0.025.xlsx

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0.0031.xlsx

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0.0063.xlsx

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0.0125.xlsx

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0mM.xlsx

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0.1mM.xlsx

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0.05mM_ADP_MAPP.xlsx

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0.0016mM_ADP.xlsx

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0.025mM_ADP.xlsx

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0.0031mM_ADP.xlsx

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0.0063mM_ADP.xlsx

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0.0125mM_ADP.xlsx

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0mM_ADP.xlsx

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Initial_rates_1ng_tAK.xlsx

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Initial_rates_5ng_tAK.xlsx

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Initial_rates_10ng_tAK.xlsx

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Initial_rates_20ng_tAK.xlsx

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Initial_rates_40ng_tAK.xlsx

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Initial_Rates_-6-_-9.xlsx

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Initial_Rates_10-14.xlsx

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Standard_Curve_Data_Endpoints.xlsx

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50ul_luc.,__100ul_1x10-8M_ATP,_Vary_AMP.xlsx

data 6640 2 16748 document 6640 IR-_PP-STRIP_+_PP-MA-STRIP.xlsx application/vnd.openxmlformats-officedocument.spreadsheetml.sheet 7562bf3ca5d5437df151b90d33447617 MD5 164389 2017-06-16 13:03:28 https://researchdata.bath.ac.uk/334/30/IR-_PP-STRIP_%2B_PP-MA-STRIP.xlsx 334 30 30 application/vnd.openxmlformats-officedocument.spreadsheetml.sheet other IR spectra raw data showing polypropylene with and without maleic anhydride deposition en public

IR-_PP-STRIP_+_PP-MA-STRIP.xlsx

data archive 4498 disk0/00/00/03/34 2017-06-26 15:59:26 2024-07-17 09:21:09 2017-06-26 15:59:26 data_collection show Hathaway Hollie H.Hathaway@bath.ac.uk 0000-0002-0026-3340 University of Bath TRUE Supervisor Jenkins Toby A.T.A.Jenkins@bath.ac.uk 0000-0002-8981-3029 University of Bath Supervisor Sutton Mark 0000-0002-2288-0446 Public Health England Study into the Kinetic Properties and Surface Attachment of a Thermostable Adenylate Kinase BY0050 CA0040 GV0030 dept_chem Abstract: A thermostable adenylate kinase (tAK) has been used as model protein contaminant on surfaces, so used because residual protein after high temperature wash steps can be detected at extremely low concentrations. This gives the potential for accurate, quantitative measurement of the effectiveness of different wash processes in removing protein contamination. Current methods utilise non-covalent (physisorbtion) of tAK to surfaces, but this can be relatively easily removed. In this study, the covalent binding of tAK to surfaces was studied to provide an alternative model for surface contamination. Kinetic analysis showed that the efficiency of the enzyme expressed as the catalytic rate over the Michaelis constant (kcat/KM) increased from 8.45 ± 3.04 mM-1 s-1 in solution to 32.23 ± 3.20 or 24.46 ± 4.41 mM-1 s-1 when the enzyme was immobilised onto polypropylene or plasma activated polypropylene respectively. Maleic anhydride plasma activated polypropylene showed potential to provide a more robust challenge for washing processes as it retained significantly higher amounts of tAK enzyme than polypropylene in simple washing experiments. Inhibition of the coupled enzyme (luciferase/luciferin) system used for the detection of adenylate kinase activity, was observed for a secondary product of the reaction. This needs to be taken into consideration when using the assay to estimate cleaning efficacy. 2017 University of Bath public RightsHolder University of Bath Biotechnology and Biological Sciences Research Council https://doi.org/10.13039/501100000268 BB/K011995/1 Study of nucleotide monophosphate kinases attachment to surfaces for new sensor development Public Health England https://doi.org/10.13039/501100002141 iCASE PhD Studentship UV-Vis Luminescence spectroscopy. IR spectroscopy. Data worked up in Microsoft Excel and presented in Origin Graphing Software. https://doi.org/10.1016/j.bbrep.2015.03.011 2014 2015 en 1 10.15125/BATH-00334 https://doi.org/10.1016/j.bbrep.2015.03.011 pub open