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Published 2011 | public
Book Section - Chapter

Overexpression and Purification of the Particulate Methane Monooxygenase from Methylococcus capsulatus (Bath)

Abstract

The particulate methane monooxygenase (pMMO) is a multi-copper enzyme that mediates the facile conversion of methane to methanol in methanotrophic bacteria. As a membrane-bound multi-subunit metalloprotein, the highly active protein has been difficult to isolate and purify to homogeneity for biochemical and biophysical studies. In this chapter, we describe a method to overexpress pMMO with good specific activity in high yields in the intracytoplasmic membranes of the host organism, together with two protocols to isolate and purify the enzyme from pMMO-enriched membranes without loss of the copper cofactors and enzymatic activity.

Additional Information

© 2011 Elsevier Inc. Available online 17 March 2011. This work was supported by Academia Sinica and grants from the National Science Council of the Republic of China (NSC 95-2113-M-001-046, 97-2113-M-001-027, 98-2113-M-001-026 to S. I. C; and 97-2113-M-001-006-MY3 to S. S.-F. Yu).

Additional details

Created:
August 19, 2023
Modified:
January 13, 2024