Investigating the Biochemical and Catalytic Properties of Nitrile Hydratases

<p>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 Fe<super>3+</super> or a non-corrin Co <super>3+</super> metal ion at its active site and consist of two non-homologous subunits, &alpha; and &beta;, which form an (&alpha;&beta;)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 <italic>Comamonas testosteroni </italic> Ni1 and the Co-type NHases from <italic>Pseudonocardia thermophila</italic> JCM 3095 and <italic>Monosiga brevicollis</italic>.</p>

Name thesis:degree_name

Doctor of Philosophy (PhD)

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Chemistry

Year dc:date.available

2014

Author dc:creator

• Martinez, Salette

Contributors dc:contributor

• Chemistry
• Copyright © 2014 Salette Martinez
• Doctor of Philosophy (PhD)

• Chemistry

Repository record dc:identifier

https://ecommons.luc.edu/luc_diss/1101

OAI identifier oai:identifier

oai:ecommons.luc.edu:luc_diss-2100