THE ISOLATION AND CHARACTERIZATION OF METHANOMONAS METHANOOXIDANS BROWN AND STRAWINSKI

Abstract

Procedures for the isolation and characterization of Metkanomonas methanooxidans Brown and Strawinski are described. Isolates from varied sources are alike in cellular morphology, inasmuch as they form only microcolonies, and in their dependence on methane or methanol as carbon and energy sources for growth. Both organic and inorganic nitrogen sources are used. The organism is a Gram negative non-sporeforming rod, 1.5 to 3.0 μ by 1.0 μ in size, and motile by means of a single polar flagellum. In growing cultures the oxygen/methane ratio was approximately 1.1 and in resting cells 1.7. The R.Q. for methane with resting cells was 0.43. Resting cells were unable to oxidize organic compounds other than methane, methanol, formaldehyde, and formate. Formic acid was detected in test solutions after cell suspensions had metabolized methane, methanol, and formaldehyde. Using sodium sulphite as trapping agent for formaldehyde, it was found that 60 to 70% of the methane or methanol consumed was converted to formaldehyde. In the presence of iodoacetate, 70% of the methane consumed was present terminally as methanol. Thus it was shown that methanol, formaldehyde, and formic acid are sequential intermediates in the oxidation of methane by these organisms.