Abstract
The particulate methane monooxygenase (pMMO) from Methylosinus trichosporium OB3b was partiallypurified and characterized by measuring the effects of reducing agents and additives, and the stability ofpMMO was studied. Duroquinol was a suitable reducing agent, and pMMO was stabilized by bovine serumalbumin (BSA). Among the additivies, the copper (II) ion stimulated pMMO at low concentration andinhibited at high concentration. The optimum conditions for pMMO activity were as follows: 45 ° C, pH 6.5and 55 mM 3-morpholinopropanesulfonic acid (MOPS) buffer, and the rate of propene epoxide formationwas 13.6 nmol min mg protein. ESR spectra indicate that the copper cluster in the membrane fraction isreduced by duroquinol and oxidized by dioxygen. The result suggests that the copper cluster is containedin the active site of pMMO.
Similar content being viewed by others
References
Burrows KJ, Harrison DEF, Higgins IJ. 1984 Substrate specificities of the soluble and particulate methane mono-oxygenase of Methylosinus trichosporium OB3b. J Gen Microbiol 130, 3327-3333.
Fox BG, Froland WA, Jollie DR, Lipscomb JD. 1990 Methane monooxygenase from Methylosinus trichosporium OB3b. Methods Enzymol 188, 191-202.
Hou CT, Patel RT, Laskin AI, Barnabe N. 1980 Microbial oxidation of gaseous hydrocarbons: Oxidation of lower N-alkenes and N-alkanes by resting cell suspensions of various methylotrophic bacteria, and the effect of methane metabolites. FEMS Microbiol Lett 9, 267-270.
Juliette LY, Hyman MR, Arp DJ. 1995 Roles of bovine serum albumin and copper in the assay and stability of ammonia monooxygenase activity in vitro. J Bacteriol 177, 4908-4913.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951 Protein measurement with the folin phenol reagent. J Biol Chem 193, 265-275.
Nguyen HHT, Shiemke AK, Jacobs SJ, et al. 1994 The nature of the copper ions in the membranes containing the particulate methane monooxygenage from Methylococcus capsulatus (Bath). J Biol Chem 269, 14995-15005.
Scott D, Brannan J, Higgins IJ. 1991 The effect of growth conditions on intracytoplasmic membranes and meethane mono-oxygenase activities in Methylosinus trichosporium OB3b. J Gen Microbiol 125, 63-72.
Shiemke AK, Cook SA, Miley T, Singleton P. 1995 Detergent solubilization of membrane-bound methane monooxygenase requires plastoquinol analogs as electron donor. Arch Biochem Biophys 321, 421-428.
Zahn JA, DiSpirito AA. 1996 Membrane-associated methane monooxygenase from Methylococcus capsulatus (Bath). J Bacteriol 178, 1018-1029.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Takeguchi, M., Miyakawa, K. & Okura, I. Properties of the membranes containing the particulate methane monooxygenase from Methylosinus trichosporium OB3b. Biometals 11, 229–234 (1998). https://doi.org/10.1023/A:1009278216452
Issue Date:
DOI: https://doi.org/10.1023/A:1009278216452