Nicotinic hydroxylation

Nicotinic acid and related compounds react with l-chloro-2,4-dinitrobenzene in the manner of the cyanogen bromide reaction to yield derivative I, which possibly also decarboxylates at elevated temperature. In alkaline medium this derivative first adds an hydroxyl ion and then undergoes ring opening to yield the colored derivative II. 

The aerobic degradation of nicotine produces an A-methylpyrrolidine as the first metabolite by dehydrogenation. This is then hydroxylated at the benzylic carbon atom by an FAD-containing oxidase (Dai et al. 1968), and the y-A-methylaminobutyrate that is produced by fission of the A-methylpyrolidine ring is demethylated by an oxidase to 4-aminobutyrate (Chiribau et al. 2004). 

The degradation of nicotine has been examined extensively in Arthrobacternicotinovorans (oxydans). In strain P34, the first metabolite was 6-hydroxynicotine, and experiments with 62 and H2 0 showed that the oxygen in the hydroxyl group was derived from H2O (Hochstein and Dalton 1965). 

Hochstein LI, BP Dalton (1965) The hydroxylation of nicotine the origin of the hydroxyl oxygen. Biochem Biophys Res Commun 21 644-648. 

The degradation of nicotine has been examined extensively in Arthrobacter nicotinovorans (oxydans) in which it is mediated by a plasmid (Brandsch et al. 1982 Schenk et al. 1998). In strain P34, the hrst metabolite was 6-hydroxynicotine, and experiments with 02 and H2 0 showed that the oxygen in the hydroxyl group was derived from H2O (Hochstein and Dalton 1965). Nicotine dehydrogenase has a molecular mass of 120,000 and contains FAD, Mo, Fe, and acid-labile sulfur (Freudenberg et al. 1988). Degradation involves a series of reactions ... 

As for the aerobic degradation of pyridines, hydroxylation of the heterocyclic ring is a key reaction in the anaerobic degradation of azaarenes by Clostridia. Whereas in Clostridium barkeri, the end products are carboxylic acids, CO2, and ammonium, the anaerobic sulfate-reducing Desulfococcus niacinii degraded nicotinate completely to CO2 (Imhoff-Stuckle and Pfennig 1983), although the details of the pathway remain incompletely resolved. 

Stadtman 1971 Kung et al. 1971). Degradation is initiated by hydroxylation of the ring, and the level of nicotinic acid hydroxylase is snbstantially increased by the addition of selenite to the medinm (Imhoff and Andreesen 1979). Nicotinate hydroxylase from Clostridium barkeri contains molybdenum that is coordinated to seleninm, which is essential for hydroxylase activity (Gladyshev et al. 1994). The most remarkable featnre of the pathway is the mechanism whereby 2-methylene-glntarate is converted into methylitaconate by a coenzyme Bi2-mediated reaction (Knng and Stadtman 1971). 

Hirschberg R, JC Ensign (1971a) Oxidation of nicotinic acid by a Bacillus species source of oxygen atoms for hydroxylation of nicotinic acid and 6-hydroxynicotinic acid. J Bacterial 108 757-759. 

Tinschert A, A Kiener, K Heinzmann, and A Tschech (1997) Isolation of new 6-methylnicotinic-acid-degrad-ing bacteria, one of which catalyses the regioselective hydroxylation of nicotinic acid at position Cj. Arch Microbiol 168 355-361. 

The B-group vitamin, nicotinic acid (259), was irradiated with low-intensity light at 254 nm. In aqueous solution without buffer, the bi-aryl (260) was obtained, presumably via decarboxylation to give the pyridyl anion which would attack position 6 of nicotinic acid. In aqueous acid, the substrate was photo-hydroxylated to give 2-hydroxynicotinic acid (40%). Clearly, only the cationic form was sufficiently activated for position 2 to be attacked by the solvent. Nicotinamide under the same conditions was also converted to the 2-hydroxy derivative, but the reaction was slower [161]. 

The first biochemical analysis of a selenium-containing XDH was reported in 1999 by Andreesen s group. This preparation was specific for xanthine and did not hydroxylate nicotinic acid. Moreover, the enzyme contained FAD, acid-labile sulfur, iron, and a dinucleotide molybdenum cofactor. Most intriguing was the near-equimolar presence of tungsten and molybdenum. It should be noted that the culture medium contained nearly equimolar levels of these metals, making one wonder whether the specificity of this enzyme for metal may be relaxed (i.e., can use Mo or W). Selenium was also found in the preparation and could be released by treatment with cyanide indicating it was also a labile cofactor. This further confirmed the chemical nature of the cofactor from the NAH enzyme from the same strain. ... 

Mouse CYP2A5 is a comparable isoform to human CYP2A6 (Raunio et al. 1988), because these isozymes are responsible for the majority of nicotine s C-oxidation (Raunio et al. 2008), cotinine s subsequent oxidation to 3 -hydroxycotinine (Sin and Tyndale 2007), and coumarin 7-hydroxylation (Kaipainen et al. 1984). Taken together, these findings suggest that mice may be a cost-effective animal model of human CYP2A6-mediated nicotine C-oxidation. 

Achromobacter xylosoxydans has been used to cany out the selective hydroxylation in high yield, using the enzyme which catalyses the first step in nicotinic acid degradation. The whole-cell biotransformation process has been scaled-up to 12 m, which is sufficient to produce high purity 6-hydroxynicotinic acid for the subsequent chemical reactions. The hydroxylation is oxygen requiring, so that oxygen transfer rate-limits the reaction. 

Figure 6 shows the proposed subunit assembly structure of the nicotinic acetylcholine receptor channel." The inner wall of the lower half part is surrounded by hydroxyl side chains from Ser and Thr, and by carboxylates or amides from Asp, Glu, and Gin at the mouth. Furthermore, a Lys residue seems to offer ion pairing with the carboxylate at the mouth. Considering the possibly similar stabilizing effect of ether and hydroxyl groups to cations, the proposed artificial supramolecular channel could be regarded as a good model of the acetylcholine receptor channel, which selects cations over anions, but does not discriminate between alkali metals. 

Figure 6. Proposed inner wall structure of the nicotinic acetylcholine receptor-channel composite from a2pY8 subunit assembly. The channel mouth is constructed from charged amino acids and their amides such as Asp, Glu, and Gin. A Lys is located at just the inner mouth. The lower half is covered by the amino acids having hydroxyl such as Ser and Thr, while the upper half is lined up with hydrophobic residues such as Leu, Val, Ala, lie, and Phe.

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