Abstract

N-Heterocyclic aromatic compounds (NHACs) are a group of organic molecules commonly produced in manufacturing and prevalent in many commercial products. Environmental contamination by NHACs is concerning given the widely reported presence of these compounds in air, water, and soil, and their toxicity and carcinogenic properties. Several common soil bacteria, including Pseudomonas putida, Eubacterium barkeri, and Bacillus niacini, have been shown to degrade NHACs to useful metabolites making mechanistic understanding of NHAC catabolism in these bacteria of interest for bioremediation efforts. The pathway of nicotinic acid (NA), a model NHAC, degradation in P. putida and E. barkeri is well understood, but the analogous pathway in B. niacini remains largely uncharacterized. The roles of three genes—FMO, HP, and DUF—in the fifteen-gene nic cluster of the B. niacini genome upregulated in the presence of NA remain undetermined. The activity of FMO, DUF, and HP are examined through endpoint kinetic assays of all three enzymes alone and together with 2,6-dihydropyridine (DHP) and 2,3,6-trihydroxypyridine (THP) as potential substrates analyzed by LC-QTOF-MS analysis. Examination of extracted substrate and product chromatograms suggest that FMO and HP together catalyze the simultaneous hydroxylation/reduction of DHP to a reduced form of THP.

Advisor

Snider, Mark

Department

Biochemistry and Molecular Biology

Disciplines

Biochemistry, Biophysics, and Structural Biology

Keywords

flavin monooxygenase, mass spectrometry, enzymology

Publication Date

2024

Degree Granted

Bachelor of Arts

Document Type

Senior Independent Study Thesis

Available for download on Tuesday, January 01, 2075

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© Copyright 2024 Katherine Olson