title: In situ structure and activity studies of an enzyme adsorbed on spectroscopically undetectable particles.
abstract: The structural characteristics and the activity of a hyperthermophilic endoglucanase were investigated upon adsorption. Silica (hydrophilic) and Teflon (hydrophobic) surfaces were selected for the study. The materials were specially designed so that the interaction of the particles with light was negligible, and the enzyme conformation in the adsorbed state was monitored in situ. The adsorption isotherms were determined, and the adsorbed endoglucanase was studied using a number of spectroscopic techniques, enzymatic activity tests, and dynamic light scattering. Experiments were performed at pH values below, at, and above the isoelectric point of the enzyme. It was shown that the enzyme adsorbed on the hydrophobic surface of Teflon with higher affinity as compared to the hydrophilic silica nanoparticles. In all cases, adsorption was followed by (slight) changes in the secondary structure resulting in decreased beta-structural content. The changes were more profound upon adsorption on Teflon. The adsorbed enzyme remained active in the adsorbed state in spite of the structural changes induced when interacting with the surfaces.
authors.affiliation: Laboratory of Physical Chemistry and Colloid Science, Wageningen University, Dreijenplein 6, 6703 HB Wageningen, The Netherlands. sotiris.koutsopoulos@wur.nl
authors.forename: Sotirios
authors.initials: S
authors.lastname: Koutsopoulos
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Tjeerdsma
authors.forename: Johannes F T
authors.initials: JF
authors.lastname: Lieshout
authors.forename: John
authors.initials: J
authors.lastname: van der Oost
authors.forename: Willem
authors.initials: W
authors.lastname: Norde
mesh_terms: D000327:Adsorption; D002480:Cellulase / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D004789:Enzyme Activation / Q000502:physiology; D008958:Models, Molecular; D010316:Particle Size; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D002480:Cellulase