1 H, 13 C, 15 N resonance assignment of the apo form of the small, chitin-active lytic polysaccharide monooxygenase Jd L
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ARTICLE
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H, 13C, 15N resonance assignment of the apo form of the small, chitin‑active lytic polysaccharide monooxygenase JdLPMO10A from Jonesia denitrificans Idd A. Christensen1 · Vincent G. H. Eijsink2 · Finn L. Aachmann1 · Gaston Courtade1 Received: 24 August 2020 / Accepted: 12 November 2020 © Springer Nature B.V. 2020
Abstract The lytic polysaccharide monooxygenase JdLPMO10A is the N-terminal domain of the multimodular protein Jd1381. The isolated JdLPMO10A domain is one of the smallest chitin-active lytic polysaccharide monooxygenases known to date with a size of only 15.5 kDa. JdLPMO10A is a copper-dependent oxidative enzyme that depolymerizes chitin by hydroxylating the C1 carbon in the glycosidic bond. JdLPMO10A has been isotopically labeled and recombinantly expressed. Here, we report the 1H, 13C, 15N resonance assignment of JdLPMO10A. Secondary structural elements predicted based on the NMR assignment are in excellent agreement with the crystal structure of JdLPMO10A. Keywords Lytic polysaccharide monooxygenase (LPMO) · Chitin
Biological context Lytic polysaccharide monooxygenases (LPMOs) are a family of copper-dependent oxidative enzymes that cleave β(1,4)-glycosidic bonds in recalcitrant polysaccharides such as cellulose and chitin (Vaaje-Kolstad et al. 2005, 2010; Quinlan et al. 2011; Hemsworth et al. 2015; Forsberg et al. 2019). LPMOs boost the degradation of crystalline polysaccharides by creating free polysaccharide chain ends that canonical glycoside hydrolases (GHs) can act on. Understanding LPMO function is therefore of interest for the biorefinery concept and may prove instrumental to achieve cost-effective saccharification of polysaccharides needed to subsequently produce biomaterials, platform chemicals and biofuels (Beeson et al. 2015). LPMOs are classified in the Auxiliary Activity (AA) families AA9-AA11 and AA13-AA16 in the Carbohydrate Active Enzymes (CAZy) database (Levasseur et al. 2013). * Gaston Courtade [email protected] 1
NOBIPOL, Department of Biotechnology and Food Science, NTNU Norwegian University of Science and Technology, Sem Sælands vei 6/8, 7491 Trondheim, Norway
Faculty of Chemistry, Biotechnology and Food Science, NMBU Norwegian University of Life Sciences, 1432 Ås, Norway
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LPMOs are copper-dependent redox enzymes that hydroxylate scissile glycosidic bonds, thus destabilizing this bond and causing cleavage (Quinlan et al. 2011; Beeson et al. 2012; Agger et al. 2014; Chylenski et al. 2019). While cellulose-active LPMOs oxidize either the C1 or C4 position, or both, only C1 oxidation has been demonstrated for chitin-active LPMOs so far. LPMOs were initially believed to catalyze a monooxygenase, i.e. an oxygen-dependent, reaction (Vaaje-Kolstad et al. 2010), but recent findings indicate that H2O2 might be the true co-substrate of LPMOs (Bissaro et al. 2017). The reaction mechanism of LPMOs remains to be uncovered in detail and is the subject of ongoing research (Chylenski et al. 2019; Courtade et al. 2020; Jones et al. 2020). JdLPMO10A is a smal
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