Hemsworth, GR orcid.org/0000-0002-8226-1380, Ciano, L, Davies, GJ et al. (1 more author) (2018) Chapter Three - Production and spectroscopic characterization of lytic polysaccharide monooxygenases. Methods in Enzymology, 613. pp. 63-90. ISSN 0076-6879
Abstract
Lytic polysaccharide monooxygenases (LPMOs, also known as PMOs) are a recently discovered family of enzymes that play a key role in the breakdown of polysaccharide substrates. The ability of LPMOs to introduce chain breaks, using an oxidative mechanism, has particularly attracted attention as the world seeks more cost-effective and environmentally friendly ways of producing second-generation biofuels for the future. LPMOs are copper-dependent enzymes and have an unusual active site which includes the N-terminal residue of the protein in the copper coordination sphere. This N-terminal histidine side chain is also methylated in fungal enzymes, the molecular reason for which is still a debated topic. The production of these enzymes poses several challenges if we are to understand their chemical mechanisms. Here, we describe the methods that have been used in the field to produce LPMOs and provide information on the workflows that we use for our electron paramagnetic resonance (EPR) spectroscopy experiments. EPR has been a particularly powerful tool in the study of these enzymes and our objective with this chapter is to provide some helpful information for researchers for whom this technique might be daunting or theoretically difficult to access.
Metadata
Item Type: | Article |
---|---|
Authors/Creators: |
|
Editors: |
|
Keywords: | LPMO; Biofuel; Bioinorganic chemistry; EPR; Protein production |
Dates: |
|
Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Structural Molecular Biology (Leeds) |
Funding Information: | Funder Grant number BBSRC BB/N019970/1 |
Depositing User: | Symplectic Publications |
Date Deposited: | 03 Dec 2018 09:55 |
Last Modified: | 04 Feb 2019 15:25 |
Status: | Published |
Publisher: | Elsevier |
Identification Number: | 10.1016/bs.mie.2018.10.014 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:139304 |