High morpholine degradation rates and formation of cytochrome P450 during growth on different cyclic amines by newly isolated Mycobacterium sp. strain HE5

• Published in: Microbiology (Society for General Microbiology) 2000-05, Vol.146 (5), p.1091-1098
• Main Authors: Schrader, Thomas, Schuffenhauer, Grit, Sielaff, Bernhard, Andreesen, Jan R
• Format: Article
• Language: English
• Subjects: Amines - metabolism; Biodegradation of pollutants; Biodegradation, Environmental; Biological and medical sciences; Biotechnology; Carbon Monoxide - analysis; Cytochrome P-450 Enzyme Inhibitors; Cytochrome P-450 Enzyme System - biosynthesis; Environment and pollution; Ethanolamine; Ethanolamine - metabolism; Fundamental and applied biological sciences. Psychology; Glycolates - metabolism; Glycolic acid; Glyoxylates - metabolism; Glyoxylic acid; Hydrogen-Ion Concentration; Industrial applications and implications. Economical aspects; Metyrapone - pharmacology; Molecular Sequence Data; Morpholine; Morpholines - metabolism; Mycobacterium; Nontuberculous Mycobacteria - growth & development; Nontuberculous Mycobacteria - isolation & purification; Nontuberculous Mycobacteria - metabolism; Piperidine; Piperidines - metabolism; Pyrrolidine; Pyrrolidines - metabolism; Soil Microbiology; Spectrophotometry, Ultraviolet; Substrate Specificity; Succinic Acid - metabolism; Temperature; Time Factors
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/00221287-146-5-1091

Institut für Mikrobiologie, Martin-Luther-Universität Halle, Kurt-Mothes-Str. 3, D-06099 Halle, Germany 1 Author for correspondence: Jan R. Andreesen. Tel: [phone]. Fax: [phone]. e-mail: j.andreesen{at}mikrobiologie.uni-halle.de Using morpholine as sole source of carbon, nitrogen and energy, strain HE5 (DSM 44238) was isolated from forest soil. The isolated strain was identified as a member of the subgroup of fast-growing Mycobacterium species as revealed by 16S rDNA analysis. An identity of 99·4% was obtained to Mycobacterium gilvum ; however, the type strain was unable to utilize morpholine. A maximal growth rate of 0·17 h -1 was observed at a morpholine concentration of 30 mM, 30 °C and pH 7·2. The substrate was tolerated at concentrations up to 100 mM. Besides morpholine, the strain utilized pyrrolidine, piperidine and proposed intermediates in morpholine metabolism such as glycolate, glyoxylate and ethanolamine. Degradation of morpholine, piperidine and pyrrolidine by resting or permeabilized cells was strictly dependent on the presence of oxygen. Addition of the cytochrome-P450-specific inhibitor metyrapone to the growth medium resulted in a significantly decreased growth rate if these cyclic amines were used as a substrate. Carbon monoxide difference spectra of crude extracts from cells grown on these substrates compared to spectra obtained for extracts of succinate-grown cells indicated that cytochrome P450 is specifically expressed during growth on the cyclic amines. These data indicated that a cytochrome-P450-dependent monooxygenase is involved in the degradation of the three cyclic amines. Keywords: morpholine, cyclic amines, degradation, Mycobacterium , cytochrome P450 The EMBL accession number of the 16S rRNA gene of Mycobacterium sp. strain HE5 is AJ012738.