Graduate Thesis Or Dissertation

 

Assaying the activities of Thermomonospora fusca E₅ and Trichoderma reesei CBHI cellulases bound to polystyrene Pubblico Deposited

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https://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/1c18dk619

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  • In this study the enzymatic activity of adsorbed Thermomonospora fusca E₅ and Trichoderma reesei CBHI cellulases were investigated using fluorescence techniques. In particular, cellulases were allowed to contact hydrophobic polystyrene surfaces under conditions of different solution concentrations, and adsorption times. Each of these variables is known to have a potential effect on enzyme structure and activity at an interface. Enzymatic activity was measured after partial elution of the adsorbed layer with both protein-free buffer and the surfactant, dodecyltrimethylammonium bromide. For E₅ at high concentration (0.5 mg/ml), adsorbed enzyme activity decreased about 20% in increasing adsorption time from 0.25 h to 24 h. At low concentration (0.001 mg/ml), adsorbed enzyme activity decreased by one order of magnitude during a 24 h period. CBHI layers lost activity only after a sufficiently long contact time with the surface, and this effect was not strongly dependent on enzyme concentrations in solution. These findings were explained with reference to structural changes undergone by adsorbed enzyme as a function of time and available interfacial area.
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