Oxygenation of methane by methane-grown Pseudomonas methanica and Methanomonas methanooxidans.

1. Experimental conditions have been found in which small amounts of methanol (approximately 2.5mm) accumulated when washed cell suspensions of methane-grown Pseudomonas methanica and Methanomonas methanooxidans were incubated with methane+oxygen mixtures in Warburg flasks. 2. The methanol formed could be separated completely from water by fractional distillation through glass helices followed by gas chromatography using 20% polyethylene glycol 400 on a Celite 545 support. 3. By using (18)O-enriched oxygen gas the abundance of (18)O in the methanol formed from oxidation of methane was measured with a Perkin-Elmer 270 combined gas chromatograph/mass spectrometer. The results showed that the oxygen in methanol was derived exclusively from gaseous oxygen in both micro-organisms. 4. Control experiments using [(18)O]water in incubation mixtures confirmed that there was negligible incorporation of the oxygen atom from water into methanol.