Date of Award

5-2014

Level of Access

Campus-Only Thesis

Degree Name

Master of Science (MS)

Department

Chemistry

Advisor

Barbara J.W. Cole

Second Committee Member

Raymond C. Fort, Jr.

Third Committee Member

Bruce L. Jensen

Abstract

Environmental concerns about chlorinated organics have created a need for new technologies to replace chlorine in pulp bleaching. One alternative is a biological bleaching that involves the oxidative enzyme laccase. Different factors affect the catalytic efficiency of the enzyme. Previous docking experiments in our group have shown that non-reducing sugar molecules like 4-methoxyphenyl-β-D-glucopyranoside can bind to the enzyme’s active site and have the potential to inhibit the activity of the enzyme. Therefore, to see the effect of such non-reducing sugars, different sets of kinetic experiments were carried out both in the presence and absence of sugar molecules. Comparison of the kinetic parameters Km and kcat obtained by preparing Lineweaver-Burk plots of their respective data were then used as the basis for the determination of the different types of enzyme inhibition. Determinations of the kinetic parameters were done using five different substrate concentrations but with the same enzyme and inhibitor concentrations. The reactions were carried out at 20°C ± 1°C and samples for analysis were quenched using NaOH prior to analysis.

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