title: Microbial transformation of 2-amino-4-methyl-3-nitropyridine.
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.
authors.affiliation: Chemical Development, Bristol-Myers Squibb, One Squibb Drive, New Brunswick, NJ 08903, USA. thomas.tully@bms.com
authors.forename: Thomas
authors.initials: T
authors.lastname: Tully
authors.forename: Mark
authors.initials: M
authors.lastname: Liu
authors.forename: Yande
authors.initials: Y
authors.lastname: Huang
authors.forename: Qingmei
authors.initials: Q
authors.lastname: Ye
authors.forename: Ramesh N
authors.initials: RN
authors.lastname: Patel
authors.forename: Animesh
authors.initials: A
authors.lastname: Goswami
mesh_terms: D001230:Aspergillus / Q000302:isolation & purification / Q000378:metabolism; D019149:Bioreactors; D001711:Biotransformation; D020093:Cunninghamella / Q000302:isolation & purification / Q000378:metabolism*; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D006900:Hydroxylation; D008970:Molecular Weight; D010087:Oxides / Q000378:metabolism; D011725:Pyridines / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D012107:Research Design; D013303:Streptomyces antibioticus / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: C584255:2-amino-4-methyl-3-nitropyridine; D010087:Oxides; D011725:Pyridines