A C4-oxidizing Lytic Polysaccharide Monooxygenase Cleaving Both Cellulose and Cello-oligosaccharides

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2013 Dec 11

PMID 24324265

Citations 140

Authors

Trine Isaksen

Bjorge Westereng

Finn L Aachmann

Jane W Agger

Daniel Kracher

Roman Kittl

Roland Ludwig

Dietmar Haltrich

Vincent G H Eijsink

Svein J Horn

Affiliations

Soon will be listed here.

Abstract

Lignocellulosic biomass is a renewable resource that significantly can substitute fossil resources for the production of fuels, chemicals, and materials. Efficient saccharification of this biomass to fermentable sugars will be a key technology in future biorefineries. Traditionally, saccharification was thought to be accomplished by mixtures of hydrolytic enzymes. However, recently it has been shown that lytic polysaccharide monooxygenases (LPMOs) contribute to this process by catalyzing oxidative cleavage of insoluble polysaccharides utilizing a mechanism involving molecular oxygen and an electron donor. These enzymes thus represent novel tools for the saccharification of plant biomass. Most characterized LPMOs, including all reported bacterial LPMOs, form aldonic acids, i.e., products oxidized in the C1 position of the terminal sugar. Oxidation at other positions has been observed, and there has been some debate concerning the nature of this position (C4 or C6). In this study, we have characterized an LPMO from Neurospora crassa (NcLPMO9C; also known as NCU02916 and NcGH61-3). Remarkably, and in contrast to all previously characterized LPMOs, which are active only on polysaccharides, NcLPMO9C is able to cleave soluble cello-oligosaccharides as short as a tetramer, a property that allowed detailed product analysis. Using mass spectrometry and NMR, we show that the cello-oligosaccharide products released by this enzyme contain a C4 gemdiol/keto group at the nonreducing end.

Citing Articles

Functional characterization of two AA10 lytic polysaccharide monooxygenases from Cellulomonas gelida.

Turunen R, Tuveng T, Forsberg Z, Schiml V, Eijsink V, Arntzen M Protein Sci. 2025; 34(3):e70060.

PMID: 39969139 PMC: 11837042. DOI: 10.1002/pro.70060.

Transcriptional and secretome analysis of Rasamsonia emersonii lytic polysaccharide mono-oxygenases.

Raheja Y, Singh V, Kumar N, Agrawal D, Sharma G, Di Falco M Appl Microbiol Biotechnol. 2024; 108(1):444.

PMID: 39167166 PMC: 11339117. DOI: 10.1007/s00253-024-13240-0.

What are the 100 most cited fungal genera?.

Bhunjun C, Chen Y, Phukhamsakda C, Boekhout T, Groenewald J, McKenzie E Stud Mycol. 2024; 108:1-411.

PMID: 39100921 PMC: 11293126. DOI: 10.3114/sim.2024.108.01.

Random forest machine-learning algorithm classifies white- and brown-rot fungi according to the number of the genes encoding Carbohydrate-Active enZyme families.

Hasegawa N, Sugiyama M, Igarashi K Appl Environ Microbiol. 2024; 90(7):e0048224.

PMID: 38832775 PMC: 11267879. DOI: 10.1128/aem.00482-24.

The disordered C-terminal tail of fungal LPMOs from phytopathogens mediates protein dimerization and impacts plant penetration.

Tamburrini K, Kodama S, Grisel S, Haon M, Nishiuchi T, Bissaro B Proc Natl Acad Sci U S A. 2024; 121(13):e2319998121.

PMID: 38513096 PMC: 10990093. DOI: 10.1073/pnas.2319998121.

References

Soding J, Biegert A, Lupas A . The HHpred interactive server for protein homology detection and structure prediction. Nucleic Acids Res. 2005; 33(Web Server issue):W244-8. PMC: 1160169. DOI: 10.1093/nar/gki408. View

Davies G, Wilson K, Henrissat B . Nomenclature for sugar-binding subsites in glycosyl hydrolases. Biochem J. 1997; 321 ( Pt 2):557-9. PMC: 1218105. DOI: 10.1042/bj3210557. View

Cantarel B, Coutinho P, Rancurel C, Bernard T, Lombard V, Henrissat B . The Carbohydrate-Active EnZymes database (CAZy): an expert resource for Glycogenomics. Nucleic Acids Res. 2008; 37(Database issue):D233-8. PMC: 2686590. DOI: 10.1093/nar/gkn663. View

Horn S, Vaaje-Kolstad G, Westereng B, Eijsink V . Novel enzymes for the degradation of cellulose. Biotechnol Biofuels. 2012; 5(1):45. PMC: 3492096. DOI: 10.1186/1754-6834-5-45. View

Phillips C, Beeson W, Cate J, Marletta M . Cellobiose dehydrogenase and a copper-dependent polysaccharide monooxygenase potentiate cellulose degradation by Neurospora crassa. ACS Chem Biol. 2011; 6(12):1399-406. DOI: 10.1021/cb200351y. View

Beeson W, Phillips C, Cate J, Marletta M . Oxidative cleavage of cellulose by fungal copper-dependent polysaccharide monooxygenases. J Am Chem Soc. 2011; 134(2):890-2. DOI: 10.1021/ja210657t. View

Hemsworth G, Taylor E, Kim R, Gregory R, Lewis S, Turkenburg J . The copper active site of CBM33 polysaccharide oxygenases. J Am Chem Soc. 2013; 135(16):6069-77. PMC: 3636778. DOI: 10.1021/ja402106e. View

Wu M, Beckham G, Larsson A, Ishida T, Kim S, Payne C . Crystal structure and computational characterization of the lytic polysaccharide monooxygenase GH61D from the Basidiomycota fungus Phanerochaete chrysosporium. J Biol Chem. 2013; 288(18):12828-39. PMC: 3642327. DOI: 10.1074/jbc.M113.459396. View

Westereng B, Agger J, Horn S, Vaaje-Kolstad G, Aachmann F, Stenstrom Y . Efficient separation of oxidized cello-oligosaccharides generated by cellulose degrading lytic polysaccharide monooxygenases. J Chromatogr A. 2012; 1271(1):144-52. DOI: 10.1016/j.chroma.2012.11.048. View

10.

Levasseur A, Drula E, Lombard V, Coutinho P, Henrissat B . Expansion of the enzymatic repertoire of the CAZy database to integrate auxiliary redox enzymes. Biotechnol Biofuels. 2013; 6(1):41. PMC: 3620520. DOI: 10.1186/1754-6834-6-41. View

11.

Harreither W, Sygmund C, Augustin M, Narciso M, Rabinovich M, Gorton L . Catalytic properties and classification of cellobiose dehydrogenases from ascomycetes. Appl Environ Microbiol. 2011; 77(5):1804-15. PMC: 3067291. DOI: 10.1128/AEM.02052-10. View

12.

Sygmund C, Kracher D, Scheiblbrandner S, Zahma K, Felice A, Harreither W . Characterization of the two Neurospora crassa cellobiose dehydrogenases and their connection to oxidative cellulose degradation. Appl Environ Microbiol. 2012; 78(17):6161-71. PMC: 3416632. DOI: 10.1128/AEM.01503-12. View

13.

Forsberg Z, Vaaje-Kolstad G, Westereng B, Bunaes A, Stenstrom Y, Mackenzie A . Cleavage of cellulose by a CBM33 protein. Protein Sci. 2011; 20(9):1479-83. PMC: 3190143. DOI: 10.1002/pro.689. View

14.

Zamocky M, Schumann C, Sygmund C, OCallaghan J, Dobson A, Ludwig R . Cloning, sequence analysis and heterologous expression in Pichia pastoris of a gene encoding a thermostable cellobiose dehydrogenase from Myriococcum thermophilum. Protein Expr Purif. 2008; 59(2):258-65. DOI: 10.1016/j.pep.2008.02.007. View

15.

Cancilla M, Wong A, Voss L, Lebrilla C . Fragmentation reactions in the mass spectrometry analysis of neutral oligosaccharides. Anal Chem. 1999; 71(15):3206-18. DOI: 10.1021/ac9813484. View

16.

Langston J, Shaghasi T, Abbate E, Xu F, Vlasenko E, Sweeney M . Oxidoreductive cellulose depolymerization by the enzymes cellobiose dehydrogenase and glycoside hydrolase 61. Appl Environ Microbiol. 2011; 77(19):7007-15. PMC: 3187118. DOI: 10.1128/AEM.05815-11. View

17.

Quinlan R, Sweeney M, Lo Leggio L, Otten H, Poulsen J, Johansen K . Insights into the oxidative degradation of cellulose by a copper metalloenzyme that exploits biomass components. Proc Natl Acad Sci U S A. 2011; 108(37):15079-84. PMC: 3174640. DOI: 10.1073/pnas.1105776108. View

18.

Kittl R, Kracher D, Burgstaller D, Haltrich D, Ludwig R . Production of four Neurospora crassa lytic polysaccharide monooxygenases in Pichia pastoris monitored by a fluorimetric assay. Biotechnol Biofuels. 2012; 5(1):79. PMC: 3500269. DOI: 10.1186/1754-6834-5-79. View

19.

Ifuku S, Saimoto H . Chitin nanofibers: preparations, modifications, and applications. Nanoscale. 2012; 4(11):3308-3318. DOI: 10.1039/c2nr30383c. View

20.

Sali A, Blundell T . Comparative protein modelling by satisfaction of spatial restraints. J Mol Biol. 1993; 234(3):779-815. DOI: 10.1006/jmbi.1993.1626. View