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Appl Environ Microbiol, January 1998, p. 153-158, Vol. 64, No. 1
Laboratoire de Synthèse,
Received 23 June 1997/Accepted 31 August 1997
Resting Mycobacterium aurum MO1 cells were incubated
with morpholine, a waste from the chemical industry. The kinetics of biodegradation was monitored by using in situ nuclear magnetic resonance (NMR). The incubation medium was directly analyzed by 1H NMR. This technique allowed the unambiguous
identification of two intermediates of the metabolic pathway involved
in the biodegradation process, glycolate and 2-(2-aminoethoxy)acetate.
The latter compound, which was not commercially available, was
synthesized, in three steps, from 2-(2-aminoethoxy)ethanol.
Quantitative analysis of the kinetics of degradation of morpholine was
performed by integrating the signals of the different metabolites in
1H-NMR spectra. Morpholine was degraded within 10 h.
The intermediates increased during the first 10 h and finally
disappeared after 20 h incubation. Assays of degradation were also
carried out with glycolate and ethanolamine, hypothetical intermediates
of the morpholine degradation pathway. They were degraded within 4 and 8 h, respectively. Until now, no tool for direct detection of intermediates or even morpholine has been available, consequently, only
hypothetical pathways have been proposed. The approach described here
gives both qualitative and quantitative information about the metabolic
routes used in morpholine degradation by M. aurum MO1. It
could be used to investigate many biodegradative processes.
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Morpholine Degradation Pathway of Mycobacterium aurum
MO1: Direct Evidence of Intermediates by In Situ 1H
Nuclear Magnetic Resonance
*
Corresponding author. Mailing address: Laboratoire de
Synthèse, Electrosynthèse et Etude de Systèmes
à Intérêt Biologique, UMR 6504 CNRS, Université
Blaise Pascal, 63177 Aubière Cedex, France. Phone: 33 4 73 40 77 14. Fax: 33 4 73 40 77 17. E-mail: amdelort{at}chimtp.univ-bpclermont.fr.
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