Abstract
The apo-form of the 24.4 kDa AA9 family lytic polysaccharide monooxygenase TaLPMO9A from Thermoascus aurantiacus has been isotopically labeled and recombinantly expressed in Pichia pastoris. In this paper, we report the 1H, 13C, and 15N chemical shift assignments, as well as an analysis of the secondary structure of the protein based on the secondary chemical shifts.
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Aachmann FL, Eijsink VGH, Vaaje-Kolstad G (2011) 1H, 13C, 15N resonance assignment of the chitin-binding protein CBP21 from Serratia marcescens. Biomol NMR Assign 5:117–119. https://doi.org/10.1007/s12104-010-9281-2
Aachmann FL, Sørlie M, Skjåk-Bræk G et al (2012) NMR structure of a lytic polysaccharide monooxygenase provides insight into copper binding, protein dynamics, and substrate interactions. Proc Natl Acad Sci USA 109:18779–18784. https://doi.org/10.1073/pnas.1208822109
Agger JW, Isaksen T, Várnai A et al (2014) Discovery of LPMO activity on hemicelluloses shows the importance of oxidative processes in plant cell wall degradation. Proc Natl Acad Sci USA 111:6287–6292. https://doi.org/10.1073/pnas.1323629111
Bennati-Granier C, Garajova S, Champion C et al (2015) Substrate specificity and regioselectivity of fungal AA9 lytic polysaccharide monooxygenases secreted by Podospora anserina. Biotechnol Biofuels 8:90. https://doi.org/10.1186/s13068-015-0274-3
Bertini I, Pierattelli R (2004) Copper(II) proteins are amenable for NMR investigations. Pure Appl Chem 76:321–333. https://doi.org/10.1351/pac200476020321
Chylenski P, Petrović DM, Müller G et al (2017) Enzymatic degradation of sulfite-pulped softwoods and the role of LPMOs. Biotechnol Biofuels 10:1–13. https://doi.org/10.1186/s13068-017-0862-5
Courtade G, Balzer S, Forsberg Z et al (2015) 1H, 13C, 15N resonance assignment of the chitin-active lytic polysaccharide monooxygenase BlLPMO10A from Bacillus licheniformis. Biomol NMR Assign 9:207–210. https://doi.org/10.1007/s12104-014-9575-x
Courtade G, Wimmer R, Dimarogona M et al (2016a) Backbone and side-chain 1H, 13C, and 15N chemical shift assignments for the apo-form of the lytic polysaccharide monooxygenase NcLPMO9C. Biomol NMR Assign 10:277–280. https://doi.org/10.1007/s12104-016-9683-x
Courtade G, Wimmer R, Røhr ÅK et al (2016b) Interactions of a fungal lytic polysaccharide monooxygenase with β-glucan substrates and cellobiose dehydrogenase. Proc Natl Acad Sci USA 113:5922–5927. https://doi.org/10.1073/pnas.1602566113
Courtade G, Forsberg Z, Vaaje-Kolstad G et al (2017) Chemical shift assignments for the apo-form of the catalytic domain, the linker region, and the carbohydrate-binding domain of the cellulose-active lytic polysaccharide monooxygenase ScLPMO10C. Biomol NMR Assign 11:257–264. https://doi.org/10.1007/s12104-017-9759-2
Couturier M, Ladevèze S, Sulzenbacher G et al (2018) Lytic xylan oxidases from wood-decay fungi unlock biomass degradation. Nat Chem Biol 14:306–310. https://doi.org/10.1038/nchembio.2558
Forsberg Z, Vaaje-Kolstad G, Westereng B et al (2011) Cleavage of cellulose by a CBM33 protein. Protein Sci 20:1479–1483. https://doi.org/10.1002/pro.689
Frommhagen M, Sforza S, Westphal AH et al (2015) Discovery of the combined oxidative cleavage of plant xylan and cellulose by a new fungal polysaccharide monooxygenase. Biotechnol Biofuels 8:101. https://doi.org/10.1186/s13068-015-0284-1
Hemsworth GR, Henrissat B, Davies GJ, Walton PH (2014) Discovery and characterization of a new family of lytic polysaccharide monooxygenases. Nat Chem Biol 10:122–126. https://doi.org/10.1038/nchembio.1417
Hu J, Arantes V, Pribowo A (2014) Substrate factors that influence the synergistic interaction of AA9 and cellulases during the enzymatic hydrolysis of biomass. Energy Environ Sci 7:2308–2315. https://doi.org/10.1039/C4EE00891J
Isaksen T, Westereng B, Aachmann FL et al (2014) A C4-oxidizing lytic polysaccharide monooxygenase cleaving both cellulose and cello-oligosaccharides. J Biol Chem 289:2632–2642. https://doi.org/10.1074/jbc.M113.530196
Keller R (2004) The computer aided resonance assignment tutorial. CANTINA Verlag, Goldau
Levasseur A, Drula E, Lombard V et al (2013) Expansion of the enzymatic repertoire of the CAZy database to integrate auxiliary redox enzymes. Biotechnol Biofuels 6:41–54. https://doi.org/10.1186/1754-6834-6-41
Lo Leggio L, Simmons TJ, Poulsen JN et al (2015) Structure and boosting activity of a starch-degrading lytic polysaccharide monooxygenase. Nat Commun 6:1–9. https://doi.org/10.1038/ncomms6961
Marsh J, Singh VK, Jia Z, Forman-Kay JD (2006) Sensitivity of secondary structure propensities to sequence differences between alpha- and gamma-synuclein: implications for fibrillation. Protein Sci 15:2795–2804. https://doi.org/10.1110/ps.062465306
Meier KK, Jones SM, Kaper T et al (2018) Oxygen activation by Cu LPMOs in recalcitrant carbohydrate polysaccharide conversion to monomer sugars. Chem Rev 118:2593–2635. https://doi.org/10.1021/acs.chemrev.7b00421
Müller G, Várnai A, Johansen KS et al (2015) Harnessing the potential of LPMO-containing cellulase cocktails poses new demands on processing conditions. Biotechnol Biofuels 8:187. https://doi.org/10.1186/s13068-015-0376-y
Pickford AR, O’Leary JM (2004) Isotopic labeling of recombinant proteins from the methylotrophic yeast Pichia pastoris. Methods Mol Biol 278:17–33. https://doi.org/10.1385/1-59259-809-9:017
Quinlan RJ, Sweeney MD, Lo Leggio L et al (2011) Insights into the oxidative degradation of cellulose by a copper metalloenzyme that exploits biomass components. Proc Natl Acad Sci USA 108:15079–15084. https://doi.org/10.1073/pnas.1105776108
Sabbadin F, Hemsworth GR, Ciano L et al (2018) An ancient family of lytic polysaccharide monooxygenases with roles in arthropod development and biomass digestion. Nat Commun 9:756. https://doi.org/10.1038/s41467-018-03142-x
Shen Y, Bax A (2013) Protein backbone and sidechain torsion angles predicted from NMR chemical shifts using artificial neural networks. J Biomol NMR 56:227–241. https://doi.org/10.1007/s10858-013-9741-y
Vaaje-Kolstad G, Westereng B, Horn SJ et al (2010) An oxidative enzyme boosting the enzymatic conversion of recalcitrant polysaccharides. Science 330:219–222. https://doi.org/10.1126/science.1192231
Vaaje-Kolstad G, Forsberg Z, Loose JS et al (2017) Structural diversity of lytic polysaccharide monooxygenases. Curr Opin Struct Biol 44:67–76. https://doi.org/10.1016/j.sbi.2016.12.012
Várnai A, Tang C, Bengtsson O et al (2014) Expression of endoglucanases in Pichia pastoris under control of the GAP promoter. Microb Cell Fact 13:1–10. https://doi.org/10.1186/1475-2859-13-57
Vu VV, Beeson WT, Phillips CM et al (2014a) Determinants of regioselective hydroxylation in the fungal polysaccharide monooxygenases. J Am Chem Soc 2:562–565. https://doi.org/10.1021/ja409384b
Vu VV, Beeson WT, Span EA et al (2014b) A family of starch-active polysaccharide monooxygenases. Proc Natl Acad Sci USA 111:13822–13827. https://doi.org/10.1073/pnas.1408090111
Zhang H, Neal S, Wishart DS (2003) RefDB: a database of uniformly referenced protein chemical shifts. J Biomol NMR 25:173–195. https://doi.org/10.1023/A:1022836027055
Acknowledgements
This work was financed by SO-funds from NTNU Norwegian University of Science and Technology and by the “Advancing biomass technology—a biomimetic approach” project, the KIFEE project and the Norwegian NMR Platform, all from the Research Council of Norway (Grant Nos. 243663, 249797, 226244, respectively).
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Kitaoku, Y., Courtade, G., Petrović, D.M. et al. Resonance assignments for the apo-form of the cellulose-active lytic polysaccharide monooxygenase TaLPMO9A. Biomol NMR Assign 12, 357–361 (2018). https://doi.org/10.1007/s12104-018-9839-y
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DOI: https://doi.org/10.1007/s12104-018-9839-y