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Quantitative evaluation of the biosynthetic pathways leading to δ-aminolevulinic acid from the Shemin precursor glycine via the C5 pathway in Arthrobacter hyalinus by analysis of 13C-labeled coproporphyrinogen III biosynthesized from [2-13C]glycine, [1-13C]acetate, and [2-13C]acetate using 13C NMR spectroscopy

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Abstract

The biosynthetic pathways leading to δ-aminolevulinic acid (ALA) from the Shemin precursor glycine via the C5 pathway in Arthrobacter hyalinus were quantitatively evaluated by means of feeding experiments with [2-13C]glycine, sodium [1-13C]acetate, and sodium [2-13C]acetate, followed by analysis of the labeling patterns of coproporphyrinogen III (Copro’gen III) (biosynthesized from ALA) using 13C NMR spectroscopy. Two biosynthetic pathways leading to ALA from glycine via the C5 pathway were identified: i.e., transformation of glycine to l-serine catalyzed by glycine hydroxymethyltransferase, and glycine synthase-catalyzed catabolism of glycine to N 5,N 10-methylene-tetrahydrofolic acid (THF), which reacts with another molecule of glycine to afford l-serine. l-Serine is transformed to acetyl-CoA via pyruvic acid. Acetyl-CoA enters the tricarboxylic acid cycle, affording 2-oxoglutaric acid, which in turn is transformed to l-glutamic acid. The l-glutamic acid enters the C5 pathway, affording ALA in A. hyalinus. A 13C NMR spectroscopic comparison of the labeling patterns of Copro’gen III obtained after feeding of [2-13C]glycine, sodium [1-13C]acetate, and sodium [2-13C]acetate showed that [2-13C]glycine transformation and [2-13C]glycine catabolism in A. hyalinus proceed in the ratio of 52 and 48 %. The reaction of [2-13C]glycine and N 5,N 10-methylene-THF, that of glycine and N 5,N 10-[methylene-13C]methylene-THF generated from the [2-13C]glycine catabolism, and that of [2-13C]glycine and N 5,N 10-[methylene-13C]methylene-THF transformed the fed [2-13C]glycine to [1-13C]acetyl-CoA, [2-13C]acetyl-CoA, and [1,2-13C2]acetyl-CoA in the ratios of 42, 37, and 21 %, respectively. These labeled acetyl-CoAs were then incorporated into ALA. Our results provide a quantitative picture of the pathways of biosynthetic transformation to ALA from glycine in A. hyalinus.

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Acknowledgments

The author would like to thank Prof. Kajiwara for helpful discussions.

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  1. Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose-shi, Tokyo, 204-8588, Japan

    Katsumi Iida

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  1. Katsumi Iida
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Iida, K. Quantitative evaluation of the biosynthetic pathways leading to δ-aminolevulinic acid from the Shemin precursor glycine via the C5 pathway in Arthrobacter hyalinus by analysis of 13C-labeled coproporphyrinogen III biosynthesized from [2-13C]glycine, [1-13C]acetate, and [2-13C]acetate using 13C NMR spectroscopy. J Radioanal Nucl Chem 295, 1819–1827 (2013). https://doi.org/10.1007/s10967-012-2104-4

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  • DOI: https://doi.org/10.1007/s10967-012-2104-4

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