Date of Award

2014

Degree Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Chemistry

First Advisor

Chemistry

Second Advisor

Copyright © 2014 Salette Martinez

Third Advisor

Doctor of Philosophy (PhD)

Abstract

Chemical and pharmaceutical industries make extensive use of amide compounds for the manufacture of commodity chemicals (e.g., acrylamide) and drug intermediates (e.g., nicotinamide). Production of amide compounds is typically achieved by the hydration of nitrile compounds under acidic or basic conditions, high temperatures, and copper catalysts. However, the use of such chemical methods leads to the generation of unwanted by-products and toxic wastes, in addition to low product yields and high production costs. An alternative route for amide production is the use of a natural catalyst, for example nitrile hydratases (NHase, E.C. 4.2.1.84). NHase is a metalloenzyme that efficiently converts nitriles to amides at neutral pH and ambient temperatures, thus reducing production of unwanted by-products and toxic wastes. NHase contains either a non-heme Fe3+ or a non-corrin Co 3+ metal ion at its active site and consist of two non-homologous subunits, α and β, which form an (αβ)2 heterotetramer. In order to utilize NHases to their full potential, it is crucial to understand their biochemical and catalytic properties; therefore, the goal of this research project was to gain insight into these fundamental properties. A combination of molecular biology, enzyme kinetics, UV-Visible spectroscopy, X-ray crystallography, and enzyme immobilization were used to accomplish this goal. Three nitrile hydratases were examined; these are the Fe-type NHase from Comamonas testosteroni Ni1 and the Co-type NHases from Pseudonocardia thermophila JCM 3095 and Monosiga brevicollis.

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Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.

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