Chitin-Active Lytic Polysaccharide Monooxygenases

  • Gaston Courtade
  • Finn L. AachmannEmail author
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 1142)


Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that catalyze the cleavage of 1,4-glycosidic bonds various plant cell wall polysaccharides and chitin. In contrast to glycoside hydrolases, LPMOs are active on the crystalline regions of polysaccharides and thus synergize with hydrolytic enzymes. This synergism leads to an overall increase in the biomass-degradation activity of enzyme mixtures. Chitin-active LPMOs were discovered in 2010 and are currently classified in families AA10, AA11, and AA15 of the Carbohydrate-Active enZYmes database, which include LPMOs from bacteria, fungi, insects, and viruses. LPMOs have become important enzymes both industrially and scientifically and, in this chapter, we provide a brief introduction to chitin-active LPMOs including a summary of the 20+ chitin-active LPMOs that have been characterized so far. Then, we describe their structural features, catalytic mechanism, and appended carbohydrate modules. Finally, we show how chitin-active LPMOs can be used to perform chemo-enzymatic modification of chitin substrates.


Lytic polysaccharide monooxygenase (LPMO) Copper-dependent enzymes Crystalline polysaccharides 


  1. Aachmann FL 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(46):18779–18784PubMedGoogle Scholar
  2. Agger JW 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(17):6287–6292PubMedGoogle Scholar
  3. Bacik J et al (2017) Neutron and atomic resolution X-ray structures of a lytic polysac- charide monooxygenase reveal copper-mediated dioxygen binding and evidence for N-terminal deprotonation. Biochemistry 56:2529–2532PubMedGoogle Scholar
  4. Beeson WT et al (2012) Oxidative cleavage of cellulose by fungal copper-dependent polysaccharide monooxygenases. J Am Chem Soc 134(2):890–892PubMedGoogle Scholar
  5. Bennati-Granier C et al (2015) Substrate specificity and regioselectivity of fungal AA9 lytic polysaccharide monooxygenases secreted by Podospora anserina. Biotechnol Biofuels 8(90):90–103PubMedPubMedCentralGoogle Scholar
  6. Bissaro B et al (2018) How a lytic polysaccharide monooxygenase binds crystalline chitin. Biochemistry 57(12):1893–1906PubMedGoogle Scholar
  7. Bissaro B et al (2017) Oxidative cleavage of polysaccharides by monocopper enzymes depends on H2O2. Nat Chem Biol 13(10):1123–1128PubMedGoogle Scholar
  8. Boraston AB et al (2004) Carbohydrate-binding modules: fine-tuning polysaccharide recognition. Biochem J 382:769–781PubMedPubMedCentralGoogle Scholar
  9. Borisova AS et al (2015) Structural and functional characterization of a lytic polysaccharide monooxygenase with broad substrate specificity. J Biol Chem 290(38):22955–22969PubMedPubMedCentralGoogle Scholar
  10. Breslmayr E et al (2018) A fast and sensitive activity assay for lytic polysaccharide monooxygenase. Biotechnol Biofuels 11(1)Google Scholar
  11. Brun E et al (1997) Solution structure of the cellulose-binding domain of the endoglucanase Z secreted by Erwinia chrysanthemi. Biochemistry 36(51):16074–16086PubMedGoogle Scholar
  12. Chaplin AK et al (2016) Heterogeneity in the Histidine-brace Copper Coordination Sphere in Auxiliary Activity Family 10 (AA10) Lytic Polysaccharide Monooxygenases. J Biol Chem 291(24):12838–50PubMedPubMedCentralGoogle Scholar
  13. Chiu E et al (2015) Structural basis for the enhancement of virulence by viral spindles and their in vivo crystallization. Proc Natl Acad Sci 112(13):201418798Google Scholar
  14. Courtade G et al (2015) 1H, 13C, 15N resonance assignment of the chitin-active lytic polysaccharide monooxygenase BlLPMO10A from Bacillus licheniformis. Biomol NMR Assign 9(1):207–210PubMedGoogle Scholar
  15. Courtade G et al (2018) The carbohydrate-binding module and linker of a modular lytic polysaccharide monooxygenase promote localized cellulose oxidation. J Biol Chem 293(34):13006–13015PubMedGoogle Scholar
  16. Couturier M et al (2018) Lytic xylan oxidases from wood-decay fungi unlock biomass degradation. Nat Chem Biol 14:306–310PubMedGoogle Scholar
  17. Crasson O et al (2017) Human chitotriosidase: catalytic domain or carbohydrate binding module, who’s leading HCHT’s biological function. Scientif Rep 7:2768–2777Google Scholar
  18. Crouch LI et al (2016) The contribution of non-catalytic carbohydrate binding modules to the activity lytic polysaccharide monooxygenases. J Biol Chem 291(14):7439–7449PubMedPubMedCentralGoogle Scholar
  19. Eibinger M et al (2014) Cellulose surface degradation by a lytic polysaccharide monooxygenase and its effect on cellulase hydrolytic efficiency. J Biol Chem 289(52):35929–35938PubMedPubMedCentralGoogle Scholar
  20. Fadel F et al (2016) X-Ray crystal structure of the full length human chitotriosidase (CHIT1) reveals features of its chitin binding domain. PLoS One 11(4):1–15Google Scholar
  21. Forsberg Z et al (2011) Cleavage of cellulose by a CBM33 protein. Protein Sci Publ Protein Soc 20(9):1479–1483Google Scholar
  22. Forsberg Z, Røhr AK et al (2014a) Comparative study of two chitin-active and two cellulose-active AA10-type lytic polysaccharide monooxygenases. Biochemistry 53(10):1647–1656PubMedGoogle Scholar
  23. Forsberg Z et al (2016) Structural and functional analysis of a lytic polysaccharide monooxygenase important for efficient utilization of chitin in Cellvibrio japonicus. J Biol Chem 291(14):7300–7312PubMedPubMedCentralGoogle Scholar
  24. Forsberg Z, Mackenzie AK et al (2014b) Structural and functional characterization of a conserved pair of bacterial cellulose-oxidizing lytic polysaccharide monooxygenases. Proc Natl Acad Sci USA 111(23):8446–8451PubMedGoogle Scholar
  25. Forsberg Z et al (2018) Structural determinants of bacterial lytic polysaccharide monooxygenase functionality. J Biol Chem 293(4):1397–1412PubMedGoogle Scholar
  26. Frandsen KEH et al (2016) The molecular basis of polysaccharide cleavage by lytic polysaccharide monooxygenases. Nat Chem Biol 12:298–303PubMedPubMedCentralGoogle Scholar
  27. Frandsen KEH, Lo Leggio L (2016) Lytic polysaccharide monooxygenases: a crystallographer’s view on a new class of biomass-degrading enzymes. IUCrJ 3:448–467PubMedPubMedCentralGoogle Scholar
  28. Frommhagen M et al (2015) Discovery of the combined oxidative cleavage of plant xylan and cellulose by a new fungal polysaccharide monooxygenase. Biotechnol Biofuels 8(101):101–113PubMedPubMedCentralGoogle Scholar
  29. Frommhagen M et al (2016) Lytic polysaccharide monooxygenases from Myceliophthora thermophila C1 differ in substrate preference and reducing agent specificity. Biotechnol Biofuels 9(1):186PubMedPubMedCentralGoogle Scholar
  30. Gregory RC et al (2016) Activity, stability and 3-D structure of the Cu(II) form of a chitin-active lytic polysaccharide monooxygenase from Bacillus amyloliquefaciens. Dalton Trans 45:16904–16912PubMedGoogle Scholar
  31. Gudmundsson M et al (2014) Structural and electronic snapshots during the transition from a Cu(II) to Cu(I) metal center of a lytic polysaccharide monooxygenase by X-ray photoreduction. J Biol Chem 289(27):18782–92PubMedPubMedCentralGoogle Scholar
  32. Hangasky JA, Iavarone AT, Marletta MA (2018) Reactivity of O2 versus H2O2 with polysaccharide monooxygenases. Proc Nat Acad SciGoogle Scholar
  33. Harris PV et al (2010) Stimulation of lignocellulosic biomass hydrolysis by proteins of glycoside hydrolase family 61: structure and function of a large, enigmatic family. Biochemistry 49(15):3305–3316PubMedGoogle Scholar
  34. Hemsworth GR et al (2014) Discovery and characterization of a new family of lytic polysaccharide monooxygenases. Nat Chem Biol 10(2):122–126PubMedGoogle Scholar
  35. Hemsworth GR et al (2013) The copper active site of CBM33 polysaccharide oxygenases. J Am Chem Soc 135(16):6069–6077PubMedPubMedCentralGoogle Scholar
  36. Horn SJ et al (2012) Novel enzymes for the degradation of cellulose. Biotechnol Biofuels 5:45–56PubMedPubMedCentralGoogle Scholar
  37. Hudson KL et al (2015) Carbohydrate-aromatic interactions in proteins. J Am Chem Soc 137:15152–15160PubMedPubMedCentralGoogle Scholar
  38. Hult E et al (2005) Molecular directionality in crystalline β-chitin: hydrolysis by chitinases A and B from Serratia marcescens 2170. Biochem J 388:851–856PubMedPubMedCentralGoogle Scholar
  39. Igarashi K et al (2014) Two-way traffic of glycoside hydrolase family 18 processive chitinases on crystalline chitin. Nat Commun 5:1–7Google Scholar
  40. Isaksen T et al (2014) A C4-oxidizing lytic polysaccharide monooxygenase cleaving both cellulose and cello-oligosaccharides. J Biol Chem 289(5):2632–2642PubMedGoogle Scholar
  41. Kari J et al (2014) Kinetics of cellobiohydrolase (Cel7A) variants with lowered substrate affinity. J Biol Chem 289(47):32459–32468PubMedPubMedCentralGoogle Scholar
  42. Karkehabadi S et al (2008) The first structure of a glycoside hydrolase family 61 member, Cel61B from Hypocrea jecorina, at 1.6 Å resolution. J Mol Biol 383(1):144–154Google Scholar
  43. Kojima Y et al (2016) A lytic polysaccharide monooxygenase with broad xyloglucan specificity from the brown-rot fungus Gloeophyllum trabeum and its action on cellulose-xyloglucan complexes. Appl Environ Microbiol 82(22):6557–6572PubMedPubMedCentralGoogle Scholar
  44. Kracher D et al (2016) Extracellular electron transfer systems fuel cellulose oxidative degradation. Science (New York, N.Y.), 352(6289):1098–1101Google Scholar
  45. Kruer-Zerhusen N et al (2017) Structure of a Thermobifida fusca lytic polysaccharide monooxygenase and mutagenesis of key residues. Biotechnol Biofuels 10(1):1–12Google Scholar
  46. Langston JA et al (2011) Oxidoreductive cellulose depolymerization by the enzymes cellobiose dehydrogenase and glycoside hydrolase 61. Appl Environ Microbiol 77(19):7007–7015PubMedPubMedCentralGoogle Scholar
  47. Levasseur A et al (2013) Expansion of the enzymatic repertoire of the CAZy database to integrate auxiliary redox enzymes. Biotechnol Biofuels 6:41–64PubMedPubMedCentralGoogle Scholar
  48. Loose JSM et al (2014) A rapid quantitative activity assay shows that the Vibrio cholerae colonization factor GbpA is an active lytic polysaccharide monooxygenase. FEBS Lett 588(18):3435–3440PubMedGoogle Scholar
  49. Loose JSM et al (2016) Activation of bacterial lytic polysaccharide monooxygenases with cellobiose dehydrogenase. Protein SciGoogle Scholar
  50. Loose JSM et al (2018) Multipoint precision binding of substrate protects lytic polysaccharide monooxygenases from self-destructive off-pathway processes. Biochemistry 57(28):4114–4124PubMedGoogle Scholar
  51. Lo Leggio L et al (2015) Structure and boosting activity of a starch-degrading lytic polysaccharide monooxygenase. Nat Commun 6:5961–5969PubMedPubMedCentralGoogle Scholar
  52. McLean BW et al (2002) Carbohydrate-binding modules recognize fine substructures of cellulose. J Biol Chem 277(52):50245–50254PubMedGoogle Scholar
  53. Mekasha S et al (2016) Structural and functional characterization of a small chitin-active lytic polysaccharide monooxygenase domain of a multi-modular chitinase from Jonesia denitrificans S. Ferguson. FEBS Lett 590(1):34–42Google Scholar
  54. Monreal J, Reese ET (1969) The chitinase of Serratia marcescens. Can J Microbiol 15(7):689–696PubMedGoogle Scholar
  55. Moser F et al (2008) Regulation and characterization of Thermobifida fusca carbohydrate-binding module proteins E7 and E8. Biotechnol Bioeng 100(6):1066–1077PubMedGoogle Scholar
  56. Mutahir Z et al (2018) Characterization and synergistic action of a tetra-modular lytic polysaccharide monooxygenase from Bacillus cereus. FEBS Lett 592(15):2562–2571PubMedGoogle Scholar
  57. Nakagawa YS et al (2015) A small lytic polysaccharide monooxygenase from Streptomyces griseus targeting α-and β-chitin. FEBS JGoogle Scholar
  58. Paspaliari DK et al (2015) Listeria monocytogenes has a functional chitinolytic system and an active lytic polysaccharide monooxygenase. FEBS J 282(5):921–936PubMedGoogle Scholar
  59. Peisach J, Blumberg WE (1974) Structural implications derived from the analysis of electron paramagnetic resonance spectra of natural and artificial copper proteins. Arch Biochem Biophys 165(2):691–708PubMedGoogle Scholar
  60. Petrović DM et al (2018) Methylation of the N-terminal histidine protects a lytic polysaccharide monooxygenase from auto-oxidative inactivation. Protein Sci 27:1635–1650Google Scholar
  61. Phillips CM et al (2011) Cellobiose dehydrogenase and a copper-dependent polysaccharide monooxygenase potentiate cellulose degradation by Neurospora crassa. ACS Chem Biol 6(12):1399–1406PubMedGoogle Scholar
  62. Quinlan RJ et al (2011) Insights into the oxidative degradation of cellulose by a copper metalloenzyme that exploits biomass components. Proc Natl Acad Sci USA 108(37):15079–15084PubMedGoogle Scholar
  63. Sabbadin F et al (2018) An ancient family of lytic polysaccharide monooxygenases with roles in arthropod development and biomass digestion. Nat Commun 9(756):756–767PubMedPubMedCentralGoogle Scholar
  64. Tan T-C et al (2015) Structural basis for cellobiose dehydrogenase action during oxidative cellulose degradation. Nat Commun 6(May):7542–7552PubMedPubMedCentralGoogle Scholar
  65. Vaaje-Kolstad G et al (2010) An oxidative enzyme boosting the enzymatic conversion of recalcitrant polysaccharides. Science (New York, NY), 330(6001):219–222Google Scholar
  66. Vaaje-Kolstad G et al (2012) Characterization of the chitinolytic machinery of Enterococcus faecalis V583 and high-resolution structure of its oxidative CBM33 enzyme. J Mol Biol 416(2):239–254PubMedGoogle Scholar
  67. Vaaje-Kolstad G, Houston DR et al (2005a) Crystal structure and binding properties of the Serratia marcescens chitin-binding protein CBP21. J Biol Chem 280(12):11313–11319PubMedGoogle Scholar
  68. Vaaje-Kolstad G et al (2013) The chitinolytic machinery of Serratia marcescens-a model system for enzymatic degradation of recalcitrant polysaccharides. FEBS J 280(13):3028–3049PubMedGoogle Scholar
  69. Vaaje-Kolstad G, Horn SJ et al (2005b) The non-catalytic chitin-binding protein CBP21 from Serratia marcescens is essential for chitin degradation. J Biol Chem 280(31):28492–28497PubMedGoogle Scholar
  70. Valenzuela SV et al (2017) Fast purification method of functional LPMOs from Streptomyces ambofaciens by affinity adsorption. Carbohyd Res 448:205–211Google Scholar
  71. Vermaas JV et al (2015) Effects of lytic polysaccharide monooxygenase oxidation on cellulose structure and binding of oxidized cellulose oligomers to cellulases. J Phys Chem B 119(20):6129–6143PubMedGoogle Scholar
  72. Vu VV et al (2014) A family of starch-active polysaccharide monooxygenases. Proc Natl Acad Sci USA 111(38):13822–13827PubMedGoogle Scholar
  73. Vuong TV et al (2017) Microplate-based detection of lytic polysaccharide monooxygenase activity by fluorescence-labeling of insoluble oxidized products. Biomacromol 18(2):610–616Google Scholar
  74. Walton PH, Davies GJ (2016) On the catalytic mechanisms of lytic polysaccharide monooxygenases. Curr Opin Chem Biol 31:195–207PubMedGoogle Scholar
  75. Wang D, Li J, Wong ACY et al (2018a) A colorimetric assay to rapidly determine the activities of lytic polysaccharide monooxygenases. Biotechnol Biofuels 11(1):1–11Google Scholar
  76. Wang D, Li J, Salazar-Alvarez G et al (2018b) Production of functionalised chitins assisted by fungal lytic polysaccharide monooxygenase. Green Chem 20(9):2091–2100Google Scholar
  77. Westereng B et al (2015) Enzymatic cellulose oxidation is linked to lignin by long-range electron transfer. Scientif Rep 5:18561–18577Google Scholar
  78. Wong E et al (2012) The Vibrio cholerae colonization factor GbpA possesses a modular structure that governs binding to different host surfaces. PLoS Pathog 8(1):1–12Google Scholar
  79. Xu GY et al (1995) Solution structure of a cellulose-binding domain from Cellulomonas fimi by nuclear magnetic resonance spectroscopy. Biochemistry 34(21):6993–7009PubMedGoogle Scholar

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© Springer Nature Singapore Pte Ltd. 2019

Authors and Affiliations

  1. 1.Department of Biotechnology and Food ScienceNOBIPOL, NTNU Norwegian University of Science and TechnologyTrondheimNorway

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