Open AccessSolubilisation of methane monooxygenase from Methylococcus capsulatus (Bath)
Author(s) -
SMITH D. Drummond S.,
DALTON Howard
Publication year - 1989
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1989.tb14877.x
Subject(s) - methane monooxygenase , chemistry , monooxygenase , methanotroph , enzyme , biochemistry , substrate (aquarium) , chromatography , methane , organic chemistry , biology , anaerobic oxidation of methane , cytochrome p450 , ecology
The membrane‐bound (particulate) form of methane monooxygenase from Methylococcus capsulatus (Bath) has been solubilised using the non‐ionic detergent dodecyl‐β‐ d ‐maltoside. A wide variety of detergents were tested and found to solubilise membrane proteins but did not yield methane monooxygenase in a form that could be subsequently activated. After solubilisation with dodecyl‐β‐ d ‐maltoside, enzyme activity was recovered using either egg or soya‐bean lipids. Attempts to further purify the solubilized methane monooxygenaser protein into its component polypeptides were unsuccessful and resulted in complete loss of enzyme activity. The major polypeptides present in the solubilised enzyme had molecular masses of 49 kDa, 23 kDa and 22 kDa which were similar to those seen in crude extracts [Prior, S. D. & Dalton H. (1985) J. Gen. Microbiol. 131 , 155–163]. Studies on substrate and inhibitor specificities indicated that the membrane‐associated and solubilised forms of methane monooxygenase were quite similar to each other but differed substantially from the well‐characterised soluble methane monooxygenase found in cells grown in a low copper regime and synthesised independently of the particulate methane monooxygenase.