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Cobamide distribution in Winogradsky columns supplemented with halogenated environmental contaminants
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Cobamide distribution in Winogradsky columns supplemented with halogenated environmental contaminants

Lilly Marie McQueen
Master of Science (M.S.), Drexel University
Sep 2022
DOI:
https://doi.org/10.17918/00001357
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Abstract

Chemistry Vitamin B12 Perfluorooctanoic acid Trichloroethylene Biochemistry Bioremediation Microbiology
Halogenated contaminants have been accumulating in our environment without an efficient method of remediation since their industrial use began in the 20th century. Recently, microbes that utilize cobamide-dependent reductive dehalogenases and iron-dependent reductive defluorination to degrade these compounds have been identified. Cobamides like cobalamin and pseudocobalamin are necessary cofactors for several enzymes, including a superfamily of reductive dehalogenases, and it has also been speculated that cobamides play a role in reductive defluorination. Cobamides are biosynthesized by a relatively small subset of bacteria but can be crossfed between species. To interrogate the presence and distribution of cobamides in a fresh-water environment, we determined the distribution of cobamides in a 1L Winogradsky column using liquid chromatography mass spectrometry (LCMS). Pseudocobalamin was found in the biofilm and top layers of the column and cobalamin was found throughout. A novel method of Winogradsky column construction was developed in which sediment and water from 1-liter columns was used to construct 150 mL 'miniature' Winogradsky columns, allowing for replicated testing of several experimental conditions. These columns were supplemented with trichloroethene (TCE), perfluorooctanoic acid (PFOA), and a cobamide precursor to enrich for cobamide-utilizing halogenated compound degraders in a complex microbial community. Cobamide analysis through mass spectrometry showed a variety of effects of these contaminants on cobamide identity and concentration. This experimental set up gives us a new way to study interspecies community metabolism in complex environmental samples, with the end goal of identifying microbial degraders of environmental contaminants.

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