Adsorption of thermomonospora fusca E(5) cellulase on silanized silica.
The adsorption kinetics and dodeceyltrimethylammonium-bromide-mediated elution of Thermomonospora fusca E(5) cellulase were recorded in situ, at hydrophobic, silanized silica. Experiments were performed at different solution concentrations, ranging from 0.001 to 0.70 mg/mL. Plateau values of adsorbed mass generally increased with increasing solution concentration, with the adsorbed layer being only partially eluted by buffer. Treatment with surfactant removed more of the adsorbed enzyme in each case, with the remaining adsorbed mass varying little among experiments. Adsorption of E(5) into this nonremovable state was suggested to occur early in the adsorption process and continue until some critical surface concentration was reached. Beyond this critical value of adsorbed mass, adsorption progressed with the protein adopting more loosely bound states. Adsorption kinetic data were interpreted with reference to an adsorption mechanism allowing for irreversible adsorption into two dissimilar states. These states were distinguished by differences in occupied interfacial area, and binding strength, presumably a result of differences in structure. Comparison of the data to the kinetic model based on this mechanism showed that the fraction of adsorbed molecules present in the more tightly bound state decreased as adsorption occurred from solutions of increasing concentration. However, the absolute values of more tightly bound molecules were less dependent on adsorption conditions.
Suvajittanont
W
W
Bioengineering, Oregon State University, 116 Gilmore Hall, Corvallis, Oregon 97331-3906, USA.
McGuire
J
J
Bothwell
M K
MK
D000192:Actinomycetales / Q000201:enzymology*; D000327:Adsorption; D002480:Cellulase / Q000302:isolation & purification*; D007700:Kinetics; D008954:Models, Biological; D012821:Silanes; D012822:Silicon Dioxide; D012996:Solutions; D013499:Surface Properties
D012821:Silanes; D012996:Solutions; D012822:Silicon Dioxide; C402807:endocellulase E5; D002480:Cellulase