Genetic and Functional Characterization of the Enzymes of Nicotinic Acid Degradation in Bacillus niacini

N‐ Heteroaromatic compounds (NHACs) are byproducts of fuel processing and are common components of herbicides, pesticides and pharmaceuticals that have been identified as environmental contaminants. A current approach to removing NHAC contaminants is bioremediation, which involves the use of bacteria and their degradation systems. Optimizing bioremediation efforts requires the understanding of different degradation strategies that have evolved in bacteria. The microbial degradation of nicotinic acid serves as a model system, the details of which can be translated to improving bioremediation of NHACs. Analysis of the recently sequenced genome of Bacillus niacini has putatively identified two gene clusters that collectively code for a distinct degradation pathway of nicotinic acid. This study describes the genetic characterization of key enzymes involved in the B. niacini pathway following the initial hydroxylation steps of the pyridine ring through gene knockouts and complementation studies. Further, reaction assays of the enzymes catalyzing the decarboxylation‐hydroxylation and the tripyridol ring‐opening steps will be established to confirm reaction substrates and products and to provide the platform for future mechanistic studies of these novel reactions. Support or Funding Information This work was supported by the Department of Chemistry and the Henry J. Copeland Fund for Independent Study at The College of Wooster.