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

Chan, Sunney I. and Nguyen, H.-Hoa T. and Chen, Kelvin H.-C. and Yu, Steve S.-F. (2011) Overexpression and Purification of the Particulate Methane Monooxygenase from Methylococcus capsulatus (Bath). In: Methods in Methane Metabolism. Part B, Methanotrophy. Methods in Enzymology . No.495. Elsevier , Amsterdam, pp. 177-193. ISBN 9780123869050. https://resolver.caltech.edu/CaltechAUTHORS:20110603-135022605

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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.


Item Type:Book Section
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/B978-0-12-386905-0.00012-7DOIArticle
http://www.sciencedirect.com/science/article/pii/B9780123869050000127PublisherArticle
ORCID:
AuthorORCID
Chan, Sunney I.0000-0002-5348-2723
Yu, Steve S.-F.0000-0002-3462-065X
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).
Funders:
Funding AgencyGrant Number
Academia SinicaUNSPECIFIED
National Science Council (Taipei)95-2113-M-001-046
National Science Council (Taipei)97-2113-M-001-027
National Science Council (Taipei)98-2113-M-001-026
National Science Council (Taipei)97-2113-M-001-006-MY3
Series Name:Methods in Enzymology
Issue or Number:495
Record Number:CaltechAUTHORS:20110603-135022605
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20110603-135022605
Official Citation:Sunney I. Chan, H.-Hoa T. Nguyen, Kelvin H.-C. Chen, Steve S.-F. Yu, Overexpression and Purification of the Particulate Methane Monooxygenase from Methylococcus capsulatus (Bath), In: Amy C. Rosenzweig and Stephen W. Ragsdale, Editor(s), Methods in Enzymology, Academic Press, 2011, Volume 495, Methods in Methane Metabolism, Part B: Methanotrophy, Pages 177-193, ISSN 0076-6879, ISBN 9780123869050, DOI: 10.1016/B978-0-12-386905-0.00012-7. (http://www.sciencedirect.com/science/article/pii/B9780123869050000127)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:23901
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:21 Jun 2011 16:51
Last Modified:09 Mar 2020 13:19

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