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Hydrophobic Effects on Cellulase Binding

EPA Grant Number: U915232
Title: Hydrophobic Effects on Cellulase Binding
Investigators: Baker, Carolyn S.
Institution: Oregon State University
EPA Project Officer: Thompson, Delores
Project Period: January 1, 1997 through January 1, 2000
Project Amount: $102,000
RFA: STAR Graduate Fellowships (1997)
Research Category: Academic Fellowships , Fellowship - Agricultural Engineering , Engineering and Environmental Chemistry

Description:

Objective:

The overall objective of this research project is to study cellulase behavior upon adsorption to hydrophobic and hydrophilic surfaces. The specific objectives of this research project are to: (1) determine if Trichoderma reesei CBH I and Thermomonospora fusca E5 cellulases adsorb to hydrophilic and hydrophobic surfaces; (2) determine if activity is lost upon adsorption to these surfaces; and (3) determine if CHBI and E5 undergo conformational charges upon adsorption, and if so, measure the kinetics of the changes.

Approach:

Colloidal polystyrene microspheres and silica nanoparticles are being used as model hydrophobic and hydrophilic surfaces, respectively. Colloidal particles such as these allow the use of solution techniques, such as size exclusion chromatography and circular dichroism spectroscopy (CD), to study protein binding and conformation. Size exclusion chromatography is being used to determine if the cellullases adsorb on hydrophilic and hydrophobic surfaces. Cellulases are mixed with particles in solution and allowed to adsorb. This cellulase-particle suspension is then passed through a gel filtration column. If the cellulase is not separated from the particles by the column, then the cellulase has adsorbed to the particle. Activity studies are being used to determine if activity is lost upon adsorption. CMCase assay with BMCC is being used to determine activity of E5 and E5-polystyrene complex. CD spectroscopy will be used to determine secondary structural changes upon adsorption.

Supplemental Keywords:

fellowship, adsorption, hydrophobic, hydrophilic, Trichoderma reesei CHB I, celluase behavior, protein binding/conformation.

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The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.

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