Abstract
Biotransformation of the highly substituted pyridine derivative 2-amino-4-methyl-3-nitropyridine by Cunninghamella elegans ATCC 26269 yielded three products each with a molecular weight of 169 Da which were identified as 2-amino-5-hydroxy-4-methyl-3-nitropyridine, 2-amino-4-hydroxymethyl-3-nitropyridine, and 2-amino-4-methyl-3-nitropyridine-1-oxide. Biotransformation by Streptomyces antibioticus ATCC 14890 gave two different products each with a molecular weight of 169 Da; one was acid labile and converted to the other stable product under acidic conditions. The structure of the stable product was established as 2-amino-4-methyl-3-nitro-6(1H)-pyridinone, and that of the less stable product was assigned as its tautomer 2-amino-6-hydroxy-4-methyl-3-nitropyridine. Four of the five biotransformation products are new compounds. Several strains of Aspergillus also converted the same substrate to the lactam 2-amino-4-methyl-3-nitro-6(1H)-pyridinone. Microbial hydroxylation by C. elegans was found to be inhibited by sulfate ion. In order to improve the yield and productivity of the 5-hydroxylation reaction by C. elegans, critical process parameters were determined and Design of Experiments (DOE) analyses were performed. Biotransformation by C. elegans was scaled up to 15-l fermentors providing 2-amino-5-hydroxy-4-methyl-3-nitropyridine at ca. 13 % yield in multi-gram levels. A simple isolation process not requiring chromatography was developed to provide purified 2-amino-5-hydroxy-4-methyl-3-nitropyridine of excellent quality.
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References
Archelas A, Furstoss R, Pohl T, Waldmann H (1995) Oxidation reactions. In: Drauz K, Wladmann H (eds) Enzyme catalysis in organic synthesis. VCH, New York, pp 667–704
Box GEP, Behnken DW (1960) Some new three-level designs for the study of quantitative variables. Technometrics 2:455–475
Box GEP, Draper NR (1987) Empirical model-building and response surfaces. Wiley, New York
Box GEP, Hunter WG, Hunter JS (1978) Statistics for experimenters: an introduction to design, data analysis, and model building. Wiley, New York. (2nd edition: Statistics for experimenters: design, innovation and discovery, 2005)
Ishikawa T, Maeda K, Hayakawa K, Kojima T (1996) Regiospecific hydroxylation of 3-(methylaminomethyl)pyridine to 5-(methylaminomethyl)-2(1H)-pyridinone by Arthrobacter ureafaciens. J Mol Catal B 1:173–179
Kiener A, Gloeckler R, Heinzmann K (1993) Preparation of 6-oxo-1,6-dihydropyridine-2-carboxylic acid by microbial hydroxylation of pyridine-2-carboxylic acid. J Chem Soc Perkin Trans I:1201–1202
Murray PJ, Onions ST, Williams JG, Joly K (2011) Preparation of pyrido[2,3-b]pyrazine compounds for use in drug formulations for treating inflammation, respiratory disorders, and viral infections. PCT Int Appl WO11158044
Tinschert A, Tschech A, Heinzmann K, Kiener A (2000) Novel regioselective hydroxylations of pyridine carboxylic acids at position C2 and pyrazine carboxylic acids at position C3. Appl Microbiol Biotechnol 53:185–195
Tully TP, Bergum JS, Schwarz SR, Durand SC, Howell JM, Patel RN, Cino PM (2007) Improvement of sordarin production through process optimization: combining traditional approaches with DOE. J Ind Microbiol Biotechnol 34:193–202
Uchida A, Yoshida T, Ogawa M, Nagasawa T (2003) Regioselective hydroxylation of quinolinic acid, lutidinic acid and isocinchomeronic acid by resting cells of pyridine dicarboxylic acid-degrading microorganisms. Appl Microbiol Biotechnol 62:337–341
Watson GK, Houghton C, Cain RB (1974) Microbial metabolism of the pyridine ring. The hydroxylation of 4-hydroxypyridine to pyridine-3,4-diol (3,4-dihydroxypyridine) by 4-hydroxypyridine-3-hydroxylase. Biochem J 140:265–276
Zefirov NS, Modyanova LV, Ouyuntsetseg A, Piskunkova NF, Terent’ev PB, Vagrov VV, Ovcharenko VV (1993) Transformation of 3-hydroxypyridine by Pseudomonas fluorescens and Rhodococcus opacus strains. Chem Heterocycl Compd 29(6):730–731
Zefirov NS, Terent’ev PB, Modyanova LV, Dovgilevich EV (1993) Regio- and stereoselective hydroxylation of some nitrogen heterocyclic compounds by microorganisms. Indian J Chem Sect B 32B:54–57
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Tully, T., Liu, M., Huang, Y. et al. Microbial transformation of 2-amino-4-methyl-3-nitropyridine. J Ind Microbiol Biotechnol 39, 1789–1799 (2012). https://doi.org/10.1007/s10295-012-1189-7
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DOI: https://doi.org/10.1007/s10295-012-1189-7