Thiomorpholine and morpholine oxidation by a cytochrome P450 in Mycobacterium aurum MO1. Evidence of the intermediates by in situ 1H NMR.
Spectrophotometric assays of Mycobacterium aurum MO1 cells extracts gave evidence of a soluble cytochrome P450, involved in the degradative pathway of morpholine, a waste product from the chemical industry. In order to get further information, the kinetics of the biodegradation of the sulfur analogue thiomorpholine was monitored by using in situ nuclear magnetic resonance (NMR). This technique allowed the identification of two intermediates: the sulfoxide of thiomorpholine resulting from S-oxidation and thiodiglycolic acid owing to ring cleavage. The S-oxidation (S-->SO) represents one of the well-known reactions catalyzed by cytochromes P450. The inhibitory effect of metyrapone, a cytochrome P450 inhibitor, on the thiomorpholine and morpholine degradative abilities of M. aurum MO1 confirmed the involvement of a cytochrome P450. These results and the decrease of the rate of formation of the first intermediate during the morpholine degradation, 2-(2-aminoethoxy) acetate, proved the key role of the cytochrome P450 in the early events of the biodegradation, i.e., in the C-N bond cleavage.
Combourieu
B
B
Laboratoire de Synthèse, Electrosynthèse et Etude de Systèmes à Intérêt Biologique, UMR 6504 CNRS, Université Blaise Pascal, Aubière, France.
Poupin
P
P
Besse
P
P
Sancelme
M
M
Veschambre
H
H
Truffaut
N
N
Delort
A M
AM
D001673:Biodegradation, Environmental; D001711:Biotransformation; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008797:Metyrapone / Q000494:pharmacology; D009025:Morpholines / Q000378:metabolism*; D009161:Mycobacterium / Q000378:metabolism*; D010084:Oxidation-Reduction; D013053:Spectrophotometry
D065607:Cytochrome P-450 Enzyme Inhibitors; D009025:Morpholines; C028971:thiamorpholine; C037574:morpholine; D003577:Cytochrome P-450 Enzyme System; D008797:Metyrapone