Oxygen-insensitive enzymatic reduction of oximes to imines.
The reduction of oximes to imines under anaerobic and aerobic conditions was studied using (E)- and (Z)-2,4,6-trimethylacetophenone oxime, benzaldoxime and (E)-2,4,6-trimethylbenzaldoxime. Pig and human liver microsomes, pig liver mitochondria and cytosol to a minor extent catalyzed the conversion of both isomeric ketoximes to the corresponding stable imine, the (E)-isomer being the better substrate. All reactions were oxygen-insensitive and required active protein and NADH or NADPH; however, NADH was preferred as cofactor. The reconstituted liver microsomal system of a pig liver CYP2D enzyme (NADH-benzamidoxime reductase), which is known to reduce N-hydroxylated derivatives of strongly basic functional groups, such as amidoximes, is also capable of reducing oximes. As expected, the corresponding imine was detected in relevant amounts when incubating 2,4,6-trimethyl-acetophenone oxime using the reconstituted enzyme system, but reduction rates were significantly lower compared to rates obtained when incubating benzamidoxime. Steric hindrance due to the methyl groups in ortho position to the oxime functionality could be excluded as being responsible for the lower conversion rates according to results obtained in incubations of 2,4,6-trimethylbenzamidoxime. When incubating benzaldoxime, only benzoic acid could be detected as metabolite, since the aldehyde is easily oxidized during incubation procedures, whereas incubations of (E)-2,4,6-trimethylbenzaldoxime also showed the formation of the corresponding aldehyde. These results allow us to postulate that the metabolism of aldoximes like 2,4,6-trimethylbenzaldoxime most likely proceeds through enzymatic reduction of the oxime to yield the intermediate imine, which is subsequently hydrolyzed to the aldehyde and then oxidized to the corresponding benzoic acid.
Heberling
Sabine
S
Pharmazeutisches Institut, Christian-Albrechts-Universität, Kiel, Germany.
Girreser
Ulrich
U
Wolf
Stephanie
S
Clement
Bernd
B
D000332:Aerobiosis; D000693:Anaerobiosis; D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology*; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007097:Imines / Q000378:metabolism*; D066298:In Vitro Techniques; D009682:Magnetic Resonance Spectroscopy; D008862:Microsomes, Liver / Q000201:enzymology*; D008930:Mitochondria, Liver / Q000201:enzymology*; D010084:Oxidation-Reduction; D010091:Oximes / Q000378:metabolism*; D013552:Swine
D007097:Imines; D010091:Oximes; D003577:Cytochrome P-450 Enzyme System; C418330:NADH-benzamidoxime reductase