Date Available
12-7-2011
Year of Publication
2008
Degree Name
Master of Science (MS)
Document Type
Thesis
College
Engineering
Department
Biosystems and Agricultural Engineering
First Advisor
Dr. Sue E. Nokes
Abstract
In recent years, fungal and bacterial cellulases have gained popularity for the conversion of lignocellulosic material to biofuels and biochemicals. This study investigated properties of fungal (Trichoderma. reesei) and bacterial (Clostridium thermocellum) cellulases. Enzymatic hydrolysis was carried out with T. reesei using nine enzyme concentration and substrate combinations. Initial rates and extents of hydrolysis were determined from the progress curve of each combination. Inhibition occurred at the higher enzyme concentrations and higher solids concentrations. Mechanisms to explain the observed inhibition are discussed. Samples of C. thermocellum purified free cellulase after 98% hydrolysis were assayed to determine the total protein content (0.15 ± 0.08 mg/mL), the enzymatic activity (0.306 ± 0.173 IU/mL) and the cellulosome mass using the Peterson method for protein determination, the cellulase activity assay with phenol-sulfuric acid assay, and the indirect ELISA adapted for C. thermocellum cellulosomes, respectively. Issues regarding reproducibility and validity of these assays are discussed.
Recommended Citation
Abadie, Alicia Renée, "QUANTIFYING CELLULASE IN HIGH-SOLIDS ENVIRONMENTS" (2008). University of Kentucky Master's Theses. 548.
https://uknowledge.uky.edu/gradschool_theses/548
