Neutron and high-resolution room-temperature X-ray data collection from crystallized lytic polysaccharide monooxygenase
Abstract
Bacteria and fungi express lytic polysaccharide monooxgyenase (LPMO) enzymes that act in conjunction with canonical hydrolytic sugar-processing enzymes to rapidly convert polysaccharides such as chitin, cellulose and starch to single monosaccharide products. In order to gain a better understanding of the structure and oxidative mechanism of these enzymes, large crystals (1–3 mm3) of a chitin-processing LPMO from the Gram-positive soil bacterium Jonesia denitrificans were grown and screened for their ability to diffract neutrons. In addition to the collection of neutron diffraction data, which were processed to 2.1 Å resolution, a high-resolution room-temperature X-ray diffraction data set was collected and processed to 1.1 Å resolution in space group P212121. To our knowledge, this work marks the first successful neutron crystallographic experiment on an LPMO. As a result, joint X-ray/neutron refinement of the resulting data will reveal new details of the structure and mechanism of this recently discovered class of enzymes.
- Authors:
-
[3];
- Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
- Norwegian Univ. of Life Sciences (Norway)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Publication Date:
- Research Org.:
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Spallation Neutron Source (SNS)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1334453
- Grant/Contract Number:
- AC05-00OR22725
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Acta Crystallographica. Section F, Structural Biology Communications
- Additional Journal Information:
- Journal Volume: 71; Journal Issue: 11; Journal ID: ISSN 2053-230X
- Publisher:
- International Union of Crystallography
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; lytic polysaccharide monooxygenase; chitin; Jonesia denitrificans; biofuel; neutron crystallography
Citation Formats
Bacik, John -Paul, Mekasha, Sophanit, Forsberg, Zarah, Kovalevsky, Andrey, Nix, Jay C., Cuneo, Matthew J., Coates, Leighton, Vaaje-Kolstad, Gustav, Chen, Julian C. -H., Eijsink, Vincent G. H., and Unkefer, Clifford J. Neutron and high-resolution room-temperature X-ray data collection from crystallized lytic polysaccharide monooxygenase. United States: N. p., 2015.
Web. doi:10.1107/S2053230X15019743.
Bacik, John -Paul, Mekasha, Sophanit, Forsberg, Zarah, Kovalevsky, Andrey, Nix, Jay C., Cuneo, Matthew J., Coates, Leighton, Vaaje-Kolstad, Gustav, Chen, Julian C. -H., Eijsink, Vincent G. H., & Unkefer, Clifford J. Neutron and high-resolution room-temperature X-ray data collection from crystallized lytic polysaccharide monooxygenase. United States. https://doi.org/10.1107/S2053230X15019743
Bacik, John -Paul, Mekasha, Sophanit, Forsberg, Zarah, Kovalevsky, Andrey, Nix, Jay C., Cuneo, Matthew J., Coates, Leighton, Vaaje-Kolstad, Gustav, Chen, Julian C. -H., Eijsink, Vincent G. H., and Unkefer, Clifford J. Thu .
"Neutron and high-resolution room-temperature X-ray data collection from crystallized lytic polysaccharide monooxygenase". United States. https://doi.org/10.1107/S2053230X15019743. https://www.osti.gov/servlets/purl/1334453.
@article{osti_1334453,
title = {Neutron and high-resolution room-temperature X-ray data collection from crystallized lytic polysaccharide monooxygenase},
author = {Bacik, John -Paul and Mekasha, Sophanit and Forsberg, Zarah and Kovalevsky, Andrey and Nix, Jay C. and Cuneo, Matthew J. and Coates, Leighton and Vaaje-Kolstad, Gustav and Chen, Julian C. -H. and Eijsink, Vincent G. H. and Unkefer, Clifford J.},
abstractNote = {Bacteria and fungi express lytic polysaccharide monooxgyenase (LPMO) enzymes that act in conjunction with canonical hydrolytic sugar-processing enzymes to rapidly convert polysaccharides such as chitin, cellulose and starch to single monosaccharide products. In order to gain a better understanding of the structure and oxidative mechanism of these enzymes, large crystals (1–3 mm3) of a chitin-processing LPMO from the Gram-positive soil bacterium Jonesia denitrificans were grown and screened for their ability to diffract neutrons. In addition to the collection of neutron diffraction data, which were processed to 2.1 Å resolution, a high-resolution room-temperature X-ray diffraction data set was collected and processed to 1.1 Å resolution in space group P212121. To our knowledge, this work marks the first successful neutron crystallographic experiment on an LPMO. As a result, joint X-ray/neutron refinement of the resulting data will reveal new details of the structure and mechanism of this recently discovered class of enzymes.},
doi = {10.1107/S2053230X15019743},
journal = {Acta Crystallographica. Section F, Structural Biology Communications},
number = 11,
volume = 71,
place = {United States},
year = {Thu Jan 01 00:00:00 EST 2015},
month = {Thu Jan 01 00:00:00 EST 2015}
}
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Works referencing / citing this record:
Detection and Characterization of a Novel Copper‐Dependent Intermediate in a Lytic Polysaccharide Monooxygenase
journal, November 2019
- Singh, Raushan K.; Blossom, Benedikt M.; Russo, David A.
- Chemistry – A European Journal, Vol. 26, Issue 2