title: 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 mm(3)) 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. 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.affiliation: Protein Crystallography Station, Bioscience Division, Los Alamos National Laboratory, TA-03, Bldg 4200, MS T007, Los Alamos, NM 87545, USA.
authors.forename: John Paul
authors.initials: JP
authors.lastname: Bacik
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, PO Box 5003, 1430 Ås, Norway.
authors.forename: Sophanit
authors.initials: S
authors.lastname: Mekasha
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, PO Box 5003, 1430 Ås, Norway.
authors.forename: Zarah
authors.initials: Z
authors.lastname: Forsberg
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, 1 Bethel Valley Road, PO Box 2008, Oak Ridge, NM 37831, USA.
authors.forename: Andrey
authors.initials: A
authors.lastname: Kovalevsky
authors.affiliation: Advanced Light Source, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720, USA.
authors.forename: Jay C
authors.initials: JC
authors.lastname: Nix
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, 1 Bethel Valley Road, PO Box 2008, Oak Ridge, NM 37831, USA.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Cuneo
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, 1 Bethel Valley Road, PO Box 2008, Oak Ridge, NM 37831, USA.
authors.forename: Leighton
authors.initials: L
authors.lastname: Coates
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, PO Box 5003, 1430 Ås, Norway.
authors.forename: Gustav
authors.initials: G
authors.lastname: Vaaje-Kolstad
authors.affiliation: Protein Crystallography Station, Bioscience Division, Los Alamos National Laboratory, TA-03, Bldg 4200, MS T007, Los Alamos, NM 87545, USA.
authors.forename: Julian C H
authors.initials: JC
authors.lastname: Chen
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, PO Box 5003, 1430 Ås, Norway.
authors.forename: Vincent G H
authors.initials: VG
authors.lastname: Eijsink
authors.affiliation: Protein Crystallography Station, Bioscience Division, Los Alamos National Laboratory, TA-03, Bldg 4200, MS T007, Los Alamos, NM 87545, USA.
authors.forename: Clifford J
authors.initials: CJ
authors.lastname: Unkefer
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D006094:Gram-Positive Bacteria / Q000201:enzymology; D006899:Mixed Function Oxygenases / Q000737:chemistry* / Q000302:isolation & purification; D033363:Neutron Diffraction / Q000379:methods*; D011135:Polysaccharides, Bacterial / Q000737:chemistry* / Q000302:isolation & purification; D013696:Temperature
chemical_list: D011135:Polysaccharides, Bacterial; D006899:Mixed Function Oxygenases
keywords: Jonesia denitrificans; biofuel; chitin; lytic polysaccharide monooxygenase; neutron crystallography