P([O-])([O-])([O-])=O.[K+].[K+].[K+].[C:9]([C:11]1[CH:12]=[N:13][CH:14]=[CH:15][CH:16]=1)#[N:10].C(#[N:19])C.[OH2:20].CN(C)[C:23](=[O:30])[C:24]1[CH:29]=[CH:28][CH:27]=C[CH:25]=1>C(#N)C>[C:9]([NH2:10])(=[O:30])[C:11]1[CH:16]=[CH:15][CH:14]=[N:13][CH:12]=1.[C:23]([OH:30])(=[O:20])[C:24]1[CH:29]=[CH:28][CH:27]=[N:19][CH:25]=1To a 15-mL polypropylene centrifuge tube was added 3.73 mL of 50 mM potassium phosphate buffer (pH 7.0), 1.0 mL of an E. coli SW132 cell extract suspension prepared as described above, and 0.2626 g of 3-cyanopyridine. The final concentration of 3-cyanopyridine was 0.494 M. The reaction mixture was mixed on a rotating platform at 23° C. After 15 min, 7.50 mL of 95:5 acetonitrile/water containing 0.30 M N,N-dimethylbenzamide (HPLC external standard) was added to the reaction, the resulting mixture centrifuged, and a 0.100 mL of the supernatant mixed with 0.900 mL of acetonitrile and analyzed by HPLC. The conversion of 3-cyanopyridine was 100%, and the yields of nicotinamide and nicotinic acid were 100% and 0%, respectively.
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