Due to extensive use of nitroaromatic compounds in the production of dyes, pesticides, explosives, and their harmful effects on the environment, studying the enzyme mechanisms of bacteria capable of utilizing these compounds as nutrient sources have become a recent field of development. One example, the Comamonas sp. strain JS765, grows on nitrobenzene as its sole carbon, nitrogen and energy source. This bacterium contains a nitrobenzene dioxygenase enzyme system that incorporates O2 into the nitrobenzene ring to form catechol and nitrite. In this project, the genes that code for the dioxygenase enzyme were cloned and then transformed into an expression vector for over-expression in E. coli NEB 5α and E. coli BL21 (DE3). This over-expression system will facilitate purification of the normal enzyme and of mutant forms for mechanistic studies.
Davis, Paul, "Constructing a Vector for Over Expressing Nitrobenzene Dioxygenase in E. coli" (2011). Honors Student Works. 4.
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