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A Novel Way to Use Vitamin C? Characterizing L‐Ascorbate (Vitamin C) Catabolism in Ralstonia eutropha
Author(s) -
Wille Brendan,
Stack Tyler,
Carter Michael
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.651.25
Subject(s) - catabolism , biochemistry , ralstonia , mutant , bacteria , gene , enzyme , ascorbic acid , metabolic pathway , biology , chemistry , free radical scavenger , phenotype , metabolism , de novo synthesis , antioxidant , genetics , food science
L‐Ascorbate (Vitamin C) is present in the environment in plant tissues, typically at concentrations from 1–10mM, and as high as 300mM. L‐Ascorbate is also an essential radical scavenger (antioxidant) for plants and mammals and is not synthesized by bacteria. Given the ubiquity of L‐ascorbate in the environment, metabolism of L‐ascorbate necessarily contributes to ecological recycling. Ralstonia eutropha H16, a soil bacterium commonly studied for its versatile use of organic and inorganic carbon sources, can use L‐ascorbate as a carbon source via an unknown pathway. Using RNAseq, whole cell transcript levels in R. eutropha were compared between L‐ascorbate‐grown cells and succinate‐or fructose‐grown cells (controls). Putative pathway genes were identified by their increase in expression greater than 50‐fold in R. eutropha grown with L‐ascorbate. Mutant strains (lacking putative pathway genes) were generated by homologous recombination directed allelic exchange. Wild type and mutant strains were grown in liquid cultures with either L‐ascorbate or succinate as their sole carbon source and monitored by optical density to observe the phenotypic result of gene deletion. Growth studies have been performed, and L‐ascorbate no growth phenotype has been confirmed for three mutant strains. Collaborators have characterized putative pathway enzymes. Characterizing the proposed catabolic pathway of L‐ascorbate in R. eutropha will provide information regarding environmental nutrient cycling and contribute novel functions of suspected pathway enzymes to protein databases. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .