Abstract

The recently discovered lytic polysaccharide monooxygenases (LPMOs) carry out oxidative cleavage of polysaccharides and are of major importance for efficient processing of biomass. NcLPMO9C from Neurospora crassa acts both on cellulose and on non-cellulose β-glucans, including cellodextrins and xyloglucan. The crystal structure of the catalytic domain of NcLPMO9C revealed an extended, highly polar substrate-binding surface well suited to interact with a variety of sugar substrates. The ability of NcLPMO9C to act on soluble substrates was exploited to study enzyme-substrate interactions. EPR studies demonstrated that the Cu2+ center environment is altered upon substrate binding, whereas isothermal titration calorimetry studies revealed binding affinities in the low micromolar range for polymeric substrates that are due in part to the presence of a carbohydrate-binding module (CBM1). Importantly, the novel structure of NcLPMO9C enabled a comparative study, revealing that the oxidative regioselectivity of LPMO9s (C1, C4, or both) correlates with distinct structural features of the copper coordination sphere. In strictly C1-oxidizing LPMO9s, access to the solvent-facing axial coordination position is restricted by a conserved tyrosine residue, whereas access to this same position seems unrestricted in C4-oxidizing LPMO9s. LPMO9s known to produce a mixture of C1- and C4-oxidized products show an intermediate situation.

Document Type

Article

Publication Date

9-18-2015

Notes/Citation Information

Published in The Journal of Biological Chemistry, v. 290, no. 38, p. 22955-22969.

This research was originally published in The Journal of Biological Chemistry. Anna S. Borisova, Trine Isaksen, Maria Dimarogona, Abhishek A. Kognole, Geir Mathiesen, Anikó Várnai, Åsmund K. Røhr, Christina M. Payne, Morten Sørlie, Mats Sandgren, and Vincent G. H. Eijsink. Structural and Functional Characterization of a Lytic Polysaccharide Monooxygenase with Broad Substrate Specificity. The Journal of Biological Chemistry. 2015; 290:22955-22969. © the American Society for Biochemistry and Molecular Biology.

The copyright holder has granted the permission for posting the article here.

Digital Object Identifier (DOI)

https://doi.org/10.1074/jbc.M115.660183

Funding Information

This work was supported by the Faculty for Natural Resources and Agriculture, Swedish University of Agricultural Sciences through the research program MicroDrivE (Microbially Derived Energy), and by the Norwegian Research Council Grants 209335, 214613, and 216162. The authors declare that they have no conflicts of interest with the contents of this article.

Christina M. Payne is supported by the August T. Larsson Guest Researcher Programme at the Swedish University of Agricultural Sciences.

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