Improvement of the purification method for retaining the activity of the particulate methane monooxygenase from Methylosinus trichosporium OB3b |
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Authors: | Akimitsu Miyaji Toshiaki Kamachi Ichiro Okura |
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Institution: | (1) Department of Bioengineering Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama, 226-8501, Japan |
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Abstract: | The purification method of particulate methane monooxygenase (pMMO) from Methylosinus trichosporium OB3b was improved, and purified pMMO retained its activity with duroquinol as a reductant. n-Dodecyl- ,d-maltoside was used for the solubilization of pMMO and Brij 58 was used for the purification for anion exchange chromatography. Compared to the original pMMO activity in the membrane fraction, 88% of the activity was now retained in the purified material. The purified pMMO monomer (94 kDa) contained only two copper atoms and did not contain iron. Both copper ions showed only a typical type II copper EPR signal with a superhyperfine structure at the g
region, indicating that the type II copper ions play an important role as the active site of methane hydroxylation in pMMO. |
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Keywords: | Methylosinus trichosporium OB3b particulate methane monooxygenase |
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