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Assaying the activities of Thermomonospora fusca E��� and Trichoderma reesei CBHI cellulases bound to polystyrene

In this study the enzymatic activity of adsorbed Thermomonospora fusca E��� 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 E��� 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. / Graduation date: 2000

Identiferoai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/33155
Date07 October 1999
CreatorsKongruang, Sasithorn
ContributorsBothwell, Michelle K., McGuire, Joseph
Source SetsOregon State University
Languageen_US
Detected LanguageEnglish
TypeThesis/Dissertation

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