skip to main content

DOE PAGESDOE PAGES

Title: Oxygen Activation at the Active Site of a Fungal Lytic Polysaccharide Monooxygenase

Lytic polysaccharide monooxygenases have attracted vast attention owing to their abilities to disrupt glycosidic bonds via oxidation instead of hydrolysis and to enhance enzymatic digestion of recalcitrant substrates including chitin and cellulose. Here, we determined the high-resolution X-ray crystal structures of an enzyme from Neurospora crassa in the resting state and of a copper(II) dioxo intermediate complex formed in the absence of substrate. X-ray crystal structures also revealed “pre-bound” molecular oxygen adjacent to the active site. An examination of protonation states enabled by neutron crystallography and density functional theory calculations identified a role for a conserved histidine in promoting oxygen activation. Our results provide a new structural description of oxygen activation by substrate free lytic polysaccharide monooxygenases and provide insights that can be extended to reactivity in the enzyme–substrate complex.
Authors:
ORCiD logo [1] ; ORCiD logo [2] ; ORCiD logo [1]
  1. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Dept. of Molecular and Structural Biochemistry
  2. Univ. of Tennessee, Knoxville, TN (United States). Dept. of Biochemisty, Cellular and Molecular Biology; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Knoxville and Computatinal Biology Inst., Computer Science, Mathematics Division
Publication Date:
Grant/Contract Number:
AC05-00OR22725; MRI 09229719; IGERT 1069091; GM105978
Type:
Accepted Manuscript
Journal Name:
Angewandte Chemie
Additional Journal Information:
Journal Volume: 129; Journal Issue: 3; Journal ID: ISSN 0044-8249
Publisher:
German Chemical Society
Research Org:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); National Science Foundation (NSF); National Institutes of Health (NIH)
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; copper; oxidoreductases; oxygen activation; polysaccharide monooxygenases; protein structures
OSTI Identifier:
1407721

O'Dell, William B., Agarwal, Pratul K., and Meilleur, Flora. Oxygen Activation at the Active Site of a Fungal Lytic Polysaccharide Monooxygenase. United States: N. p., Web. doi:10.1002/ange.201610502.
O'Dell, William B., Agarwal, Pratul K., & Meilleur, Flora. Oxygen Activation at the Active Site of a Fungal Lytic Polysaccharide Monooxygenase. United States. doi:10.1002/ange.201610502.
O'Dell, William B., Agarwal, Pratul K., and Meilleur, Flora. 2016. "Oxygen Activation at the Active Site of a Fungal Lytic Polysaccharide Monooxygenase". United States. doi:10.1002/ange.201610502. https://www.osti.gov/servlets/purl/1407721.
@article{osti_1407721,
title = {Oxygen Activation at the Active Site of a Fungal Lytic Polysaccharide Monooxygenase},
author = {O'Dell, William B. and Agarwal, Pratul K. and Meilleur, Flora},
abstractNote = {Lytic polysaccharide monooxygenases have attracted vast attention owing to their abilities to disrupt glycosidic bonds via oxidation instead of hydrolysis and to enhance enzymatic digestion of recalcitrant substrates including chitin and cellulose. Here, we determined the high-resolution X-ray crystal structures of an enzyme from Neurospora crassa in the resting state and of a copper(II) dioxo intermediate complex formed in the absence of substrate. X-ray crystal structures also revealed “pre-bound” molecular oxygen adjacent to the active site. An examination of protonation states enabled by neutron crystallography and density functional theory calculations identified a role for a conserved histidine in promoting oxygen activation. Our results provide a new structural description of oxygen activation by substrate free lytic polysaccharide monooxygenases and provide insights that can be extended to reactivity in the enzyme–substrate complex.},
doi = {10.1002/ange.201610502},
journal = {Angewandte Chemie},
number = 3,
volume = 129,
place = {United States},
year = {2016},
month = {12}
}

Works referenced in this record:

Structural Basis for Substrate Targeting and Catalysis by Fungal Polysaccharide Monooxygenases
journal, June 2012
  • Li, Xin; Beeson, William T.; Phillips, Christopher M.
  • Structure, Vol. 20, Issue 6, p. 1051-1061
  • DOI: 10.1016/j.str.2012.04.002

Cellobiose Dehydrogenase and a Copper-Dependent Polysaccharide Monooxygenase Potentiate Cellulose Degradation by Neurospora crassa
journal, December 2011
  • Phillips, Christopher M.; Beeson, William T.; Cate, Jamie H.
  • ACS Chemical Biology, Vol. 6, Issue 12, p. 1399-1406
  • DOI: 10.1021/cb200351y

Stimulation of Lignocellulosic Biomass Hydrolysis by Proteins of Glycoside Hydrolase Family 61 Structure and Function of a Large, Enigmatic Family
journal, April 2010
  • Harris, Paul; Welner, Ditte; McFarland, K.
  • Biochemistry, Vol. 49, Issue 15, p. 3305-3316
  • DOI: 10.1021/bi100009p

An Oxidative Enzyme Boosting the Enzymatic Conversion of Recalcitrant Polysaccharides
journal, October 2010
  • Vaaje-Kolstad, Gustav; Westereng, Bjørge; Horn, Svein J.
  • Science, Vol. 330, Issue 6001, p. 219-222
  • DOI: 10.1126/science.1192231

Expansion of the enzymatic repertoire of the CAZy database to integrate auxiliary redox enzymes
journal, January 2013
  • Levasseur, Anthony; Drula, Elodie; Lombard, Vincent
  • Biotechnology for Biofuels, Vol. 6, Issue 1, Article No. 41
  • DOI: 10.1186/1754-6834-6-41