OXYMOD

• Stepnov, Anton & Eijsink, Vincent (2023). Looking at LPMO reactions through the lens of the HRP/Amplex Red assay. In Shukla, Arun K. (Eds.), Methods in Enzymology. Elsevier. ISSN 978-0-323-99264-0. p. 163–189. doi: 10.1016/bs.mie.2022.08.049.
• Votvik, Amanda Kristine; Kjendseth, Åsmund Røhr; Bissaro, Bastien; Stepnov, Anton; Sørlie, Morten & Eijsink, Vincent [Show all 7 contributors for this article] (2023). Structural and functional characterization of the catalytic domain of a cell-wall anchored bacterial lytic polysaccharide monooxygenase from Streptomyces coelicolor. Scientific Reports. ISSN 2045-2322. 13(1). doi: 10.1038/s41598-023-32263-7.
• Østby, Heidi; Tuveng, Tina Rise; Stepnov, Anton; Vaaje-Kolstad, Gustav; Forsberg, Zarah & Eijsink, Vincent (2023). Impact of Copper Saturation on Lytic Polysaccharide Monooxygenase Performance. ACS Sustainable Chemistry and Engineering. ISSN 2168-0485. doi: 10.1021/acssuschemeng.3c03714.
• Junghare, Madan; Manavalan, Tamilvendan; Fredriksen, Lasse; Leiros, Ingar; Altermark, Bjørn & Eijsink, Vincent [Show all 7 contributors for this article] (2023). Biochemical and structural characterisation of a family GH5 cellulase from endosymbiont of shipworm P. megotara. Biotechnology for Biofuels and Bioproducts. 16(1), p. 1–15. doi: 10.1186/s13068-023-02307-1. Full text in Research Archive
• Østby, Heidi; Christensen, Idd Andrea; Hennum, Karen; Varnai, Aniko; Buchinger, Edith & Grandal, Siri [Show all 10 contributors for this article] (2023). Functional characterization of a lytic polysaccharide monooxygenase from Schizophyllum commune that degrades non-crystalline substrates. Scientific Reports. ISSN 2045-2322. 13. doi: 10.1038/s41598-023-44278-1. Full text in Research Archive
• Forsberg, Zarah; Stepnov, Anton; Tesei, Giulio; Wang, Yong; Buchinger, Edith & Kristiansen, Sandra Kvarsvik [Show all 11 contributors for this article] (2023). The effect of linker conformation on performance and stability of a two-domain lytic polysaccharide monooxygenase. Journal of Biological Chemistry. ISSN 0021-9258. 299(11). doi: 10.1016/j.jbc.2023.105262. Full text in Research Archive
• Kommedal, Eirik; Angeltveit, Camilla Fløien; Klau, Leesa Jane; Ayuso-Fernandez, Ivan; Arstad, Bjørnar & Antonsen, Simen [Show all 13 contributors for this article] (2023). Visible light-exposed lignin facilitates cellulose solubilization by lytic polysaccharide monooxygenases. Nature Communications. ISSN 2041-1723. 14(1). doi: 10.1038/s41467-023-36660-4. Full text in Research Archive
• Myklebust, Alexander (2023). Innovasjon som kombinasjon, teknologi og omstilling. Norsk Filosofisk tidsskrift. ISSN 0029-1943. doi: 10.18261/nft.58.2-3.6. Full text in Research Archive
• Christensen, Idd Andrea; Eijsink, Vincent; Stepnov, Anton; Courtade, Gaston & Aachmann, Finn Lillelund (2023). Following the Fate of Lytic Polysaccharide Monooxygenases under Oxidative Conditions by NMR Spectroscopy. Biochemistry. ISSN 0006-2960. 62(12), p. 1976–1993. doi: 10.1021/acs.biochem.3c00089. Show summary

  Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that catalyze oxidative cleavage of polysaccharides, such as cellulose and chitin. LPMO catalysis requires a reductant, such as ascorbic acid, and hydrogen peroxide, which can be generated in situ in the presence of molecular oxygen and various electron donors. While it is known that reduced LPMOs are prone to autocatalytic oxidative damage due to off-pathway reactions with the oxygen co-substrate, little is known about the structural consequences of such damage. Here, we present atomic-level insights into how the structure of the chitin-active SmLPMO10A is affected by oxidative damage using NMR and circular dichroism spectroscopy. Incubation with ascorbic acid could lead to rearrangements of aromatic residues, followed by more profound structural changes near the copper-active site and loss of activity. Longer incubation times induced changes in larger parts of the structure, indicative of progressing oxidative damage. Incubation with ascorbic acid in the presence of chitin led to similar changes in the observable (i.e., not substrate-bound) fraction of the enzyme. Upon subsequent addition of H2O2, which drastically speeds up chitin hydrolysis, NMR signals corresponding to seemingly intact SmLPMO10A reappeared, indicating dissociation of catalytically competent LPMO. Activity assays confirmed that SmLPMO10A retained catalytic activity when pre-incubated with chitin before being subjected to conditions that induce oxidative damage. Overall, this study provides structural insights into the process of oxidative damage of SmLPMO10A and demonstrates the protective effect of the substrate.

• Martins, Margarida; Carvalheiro, Florbela & Gírio, Francisco (2022). An overview of lignin pathways of valorization: from isolation to Refining and conversion into value added products. Biomass Conversion and Biorefinery. ISSN 2190-6815. doi: 10.1007/s13399-022-02701-z.
• Stepnov, Anton; Eijsink, Vincent & Forsberg, Zarah (2022). Enhanced in situ H<inf>2</inf>O<inf>2</inf> production explains synergy between an LPMO with a cellulose-binding domain and a single-domain LPMO. Scientific Reports. ISSN 2045-2322. 12(1). doi: 10.1038/s41598-022-10096-0.
• Cordas, Cristina M.; Valério, Gabriel N.; Stepnov, Anton; Kommedal, Eirik; Kjendseth, Åsmund Røhr & Forsberg, Zarah [Show all 8 contributors for this article] (2022). Electrochemical characterization of a family AA10 LPMO and the impact of residues shaping the copper site on reactivity. Journal of Inorganic Biochemistry. ISSN 0162-0134. doi: 10.1016/j.jinorgbio.2022.112056.
• Myskja, Bjørn Kåre & Myklebust, Alexander (2022). Socratic dialogue on responsible innovation – a methodological experiment in empirical ethics. Etikk i praksis. ISSN 1890-3991. 17(1), p. 29–44. doi: 10.5324/eip.v17i1.4950. Full text in Research Archive
• Nguyen, Giang-Son; Lewin, Anna; di Bartolomeo, Francesca & Wentzel, Alexander (2021). Recent Advances in Enzymatic Conversion of Lignin to Value Added Products. In Moura, José J. G.; Moura, Isabel & Maia, Luisa B. (Ed.), Enzymes for Solving Humankind's Problems. Springer Publishing Company. ISSN 978-3-030-58315-6. p. 439–471. doi: 10.1007/978-3-030-58315-6_14. Show summary

  Lignin is the second most abundant biopolymer and the largest reservoir of bio-based aromatic building blocks on earth that could replace numerous products currently produced from fossil feedstock with more sustainable, bio-based alternatives. However, its large-scale exploitation for value creation beyond its current main use in energy applications is hampered by its natural inertness and structural complexity and the highly denatured state of the bulk of technical lignin available as a by-product of large-scale processing of lignocellulose in the pulp and paper industry. In this section, we outline current paths of lignin valorization for value creation with a focus on enzymatic processing. Microbial sources and the current status of knowledge of the most important classes of ligninolytic enzymes are presented, as are options for new enzyme discovery and possible approaches to solve some of the key challenges of efficient enzymatic depolymerization of lignin to derive compounds directly marketable as value-added chemicals or usable as a feedstock for further upgrading by microbial metabolism to bio-based fuels, chemicals, and materials.

• Stepnov, Anton; Christensen, Idd Andrea; Forsberg, Zarah; Aachmann, Finn Lillelund; Courtade, Gaston & Eijsink, Vincent (2021). The impact of reductants on the catalytic efficiency of a lytic polysaccharide monooxygenase and the special role of dehydroascorbic acid. FEBS Letters. ISSN 0014-5793. doi: 10.1002/1873-3468.14246.
• Madland, Eva; Forsberg, Zarah; Wang, Yong; Lindorff-Larsen, Kresten; Niebisch, Axel & Modregger, Jan [Show all 9 contributors for this article] (2021). Structural and functional variation of chitin-binding domains of a lytic polysaccharide monooxygenase from Cellvibrio japonicus. Journal of Biological Chemistry. ISSN 0021-9258. 297(4), p. 1–17. doi: 10.1016/j.jbc.2021.101084. Full text in Research Archive
• Rieder, Lukas; Stepnov, Anton; Sørlie, Morten & Eijsink, Vincent (2021). Fast and Specific Peroxygenase Reactions Catalyzed by Fungal Mono-Copper Enzymes. Biochemistry. ISSN 0006-2960. 60, p. 3633–3643. doi: 10.1021/acs.biochem.1c00407.
• Manavalan, Tamilvendan; Stepnov, Anton; Hegnar, Olav Aaseth & Eijsink, Vincent (2021). Sugar oxidoreductases and LPMOs – two sides of the same polysaccharide degradation story? Carbohydrate Research. ISSN 0008-6215. 505. doi: 10.1016/j.carres.2021.108350.
• Stepnov, Anton; Forsberg, Zarah; Sørlie, Morten; Nguyen, Giang-Son; Wentzel, Alexander & Kjendseth, Åsmund Røhr [Show all 7 contributors for this article] (2021). Unraveling the roles of the reductant and free copper ions in LPMO kinetics. Biotechnology for Biofuels. ISSN 1754-6834. 14(28). doi: 10.1186/s13068-021-01879-0. Full text in Research Archive Show summary

  Background: Lytic polysaccharide monooxygenases (LPMOs) are monocopper enzymes that catalyze oxidative depolymerization of industrially relevant crystalline polysaccharides, such as cellulose, in a reaction that depends on an electron donor and O2 or H2O2. While it is well known that LPMOs can utilize a wide variety of electron donors, the variation in reported efciencies of various LPMO-reductant combinations remains largely unexplained. Results: In this study, we describe a novel two-domain cellulose-active family AA10 LPMO from a marine actinomy‑ cete, which we have used to look more closely at the efects of the reductant and copper ions on the LPMO reaction. Our results show that ascorbate-driven LPMO reactions are extremely sensitive to very low amounts (micromolar concentrations) of free copper because reduction of free Cu(II) ions by ascorbic acid leads to formation of H2O2, which speeds up the LPMO reaction. In contrast, the use of gallic acid yields steady reactions that are almost insensitive to the presence of free copper ions. Various experiments, including dose–response studies with the enzyme, showed that under typically used reaction conditions, the rate of the reaction is limited by LPMO-independent formation of H2O2 resulting from oxidation of the reductant. Conclusion: The strong impact of low amounts of free copper on LPMO reactions with ascorbic acid and O2, i.e. the most commonly used conditions when assessing LPMO activity, likely explains reported variations in LPMO rates. The observed diferences between ascorbic acid and gallic acid show a way of making LPMO reactions less copperdependent and illustrate that reductant efects on LPMO action need to be interpreted with great caution. In clean reactions, with minimized generation of H2O2, the (O2-driven) LPMO reaction is exceedingly slow, compared to the much faster peroxygenase reaction that occurs when adding H2O2.

• Cordas, Cristina M.; Nguyen, Giang-Son; Valério, Gabriel N.; Jønsson, Malene; Söllner, Katharina & Aune, Ingvild [Show all 8 contributors for this article] (2021). Discovery and characterization of a novel Dyp-type peroxidase from a marine actinobacterium isolated from Trondheim fjord, Norway. Journal of Inorganic Biochemistry. ISSN 0162-0134. 226. doi: 10.1016/j.jinorgbio.2021.111651. Show summary

  A new dye-decolorizing peroxidase (DyP) was discovered through a data mining workflow based on HMMER software and profile Hidden Markov Model (HMM) using a dataset of 1200 genomes originated from a Actinobacteria strain collection isolated from Trondheim fjord. Instead of the conserved GXXDG motif known for Dyp-type peroxidases, the enzyme contains a new conserved motif EXXDG which has been not reported before. The enzyme can oxidize an anthraquinone dye Remazol Brilliant Blue R (Reactive Blue 19) and other phenolic compounds such as ferulic acid, sinapic acid, caffeic acid, 3-methylcatechol, dopamine hydrochloride, and tannic acid. The acidic pH optimum (3 to 4) and the low temperature optimum (25 °C) were confirmed using both biochemical and electrochemical assays. Kinetic and thermodynamic parameters associated with the catalytic redox center were attained by electrochemistry.

• Courtade, Gaston; Ciano, Luisa; Paradisi, Alessandro; Lindley, Peter J.; Forsberg, Zarah & Sørlie, Morten [Show all 11 contributors for this article] (2020). Mechanistic basis of substrate–O2 coupling within a chitin-active lytic polysaccharide monooxygenase: An integrated NMR/EPR study. Proceedings of the National Academy of Sciences of the United States of America. ISSN 0027-8424. 117(32), p. 19178–19189. doi: 10.1073/pnas.2004277117. Full text in Research Archive
• Guerrero-Garzón, Jaime Felipe; Madland, Eva; Zehl, Martin; Singh, Madhurendra; Rezaei, Shiva & Aachmann, Finn Lillelund [Show all 16 contributors for this article] (2020). Class IV lasso peptides synergistically induce proliferation of cancer cells and sensitize them to doxorubicin. iScience. ISSN 2589-0042. 23(12). doi: 10.1016/j.isci.2020.101785. Full text in Research Archive
• Christensen, Idd Andrea; Eijsink, Vincent; Aachmann, Finn Lillelund & Courtade, Gaston (2020). 1H, 13C, 15N resonance assignment of the apo form of the small, chitin-active lytic polysaccharide monooxygenase JdLPMO10Afrom Jonesia denitrificans. Biomolecular NMR Assignments. ISSN 1874-2718. 15, p. 79–84. doi: 10.1007/s12104-020-09986-z. Full text in Research Archive
• Bissaro, Bastien; Streit, Bennett; Isaksen, Ingvild; Eijsink, Vincent; Beckham, Gregg T. & DuBois, Jennifer (2020). Molecular mechanism of the chitinolytic peroxygenase reaction. Proceedings of the National Academy of Sciences of the United States of America. ISSN 0027-8424. 117(3), p. 1504–1513. doi: 10.1073/pnas.1904889117.
• Forsberg, Zarah; Stepnov, Anton; Nærdal, Guro Kruge; Klinkenberg, Geir & Eijsink, Vincent (2020). Engineering lytic polysaccharide monooxygenases (LPMOs). Methods in Enzymology. ISSN 0076-6879. 644. doi: 10.1016/bs.mie.2020.04.052. Show summary

  Lytic polysaccharide monooxygenases (LPMOs) are mono-copper enzymes that catalyze the hydroxylation of glycosidic bonds found in the most abundant and recalcitrant polysaccharides on Earth. Since their discovery in 2010, these enzymes have received extensive attention in both fundamental and applied research due to their remarkable oxidative power and synergistic interplay with hydrolytic enzymes. The harsh and unnatural conditions used in industrial enzymatic saccharification processes and the sensitivity of LPMOs for damage induced by reactive oxygen species call for enzyme engineering to develop LPMOs to become robust industrial biocatalysts. Other engineering targets include improved catalytic activity, adjusted substrate specificity and the introduction of completely new activities. Reaching these targets not only requires appropriate methods for measuring enzyme activity, but also requires in-depth knowledge of the active site and the reaction mechanism, which is yet to be achieved in the LPMO field. Here we describe what has been done in the LPMO engineering field so far. Furthermore, we address the difficulties involved in properly assessing LPMO functionality, which are due to common side reactions taking place in LPMO reactions and which complicate screening methods.

• Kont, Riin; Pihlajaniemi, Ville; Borisova, Anna S.; Aro, Nina; Marjamaa, Kaisa & Loogen, Judith [Show all 10 contributors for this article] (2019). The liquid fraction from hydrothermal pretreatment of wheat straw provides lytic polysaccharide monooxygenases with both electrons and H2O2 co-substrate. Biotechnology for Biofuels. ISSN 1754-6834. 12(235). doi: 10.1186/s13068-019-1578-5.
• Perna, Valentina; Meyer, Anne S.; Holck, Jesper; Eltis, Lindsay; Eijsink, Vincent & Agger, Jane W. (2019). Laccase-Catalyzed Oxidation of Lignin Induces Production of H2O2 . ACS Sustainable Chemistry and Engineering. ISSN 2168-0485. p. 831–841. doi: 10.1021/acssuschemeng.9b04912.
• Kuusk, Silja; Kont, Riin; Kuusk, Piret; Heering, Agnes; Sørlie, Morten & Bissaro, Bastien [Show all 8 contributors for this article] (2018). Kinetic insights into the role of the reductant in H2O2-driven degradation of chitin by a bacterial lytic polysaccharide monooxygenase. Journal of Biological Chemistry. ISSN 0021-9258. 294(5), p. 1516–1528. doi: 10.1074/jbc.RA118.006196.
• Courtade, Gaston; Forsberg, Zarah; Heggset, Ellinor Bævre; Eijsink, Vincent & Aachmann, Finn Lillelund (2018). The carbohydrate-binding module and linker of a modular lytic polysaccharide monooxygenase promote localized cellulose oxidation. Journal of Biological Chemistry. ISSN 0021-9258. 293(34), p. 13006–13015. doi: 10.1074/jbc.RA118.004269. Full text in Research Archive

View all works in Cristin

• Nguyen, Giang-Son; Le, Simone Balzer; Aune, Ingvild Haugnes; Nærdal, Ingemar; Heggeset, Tonje Marita Bjerkan & Di Bartolomeo, Francesca [Show all 12 contributors for this article] (2023). Digital bioprospecting for exploring biodiversity and discovering novel bioactive compounds and industrial enzymes.
• Nguyen, Giang-Son; Le, Simone Balzer; Aune, Ingvild Haugnes; Nærdal, Ingemar; Heggeset, Tonje Marita Bjerkan & Di Bartolomeo, Francesca [Show all 12 contributors for this article] (2023). Digital bioprospecting for exploring biodiversity and discovering novel bioactive compounds and industrial enzymes.
• Redaksjon, KBM (2023). New knowledge about the role of light in the enzymatic breakdown of plant biomass published in Nature Communications. nmbu.no. Show summary

  https://www.nmbu.no/en/faculty/kbm/news/node/46727

• Junghare, Madan; Arntzen, Magnus Øverlie; Pfeiffer, Birgit; Eijsink, Vincent; Daniel, Rolf & Altermark, Bjørn [Show all 7 contributors for this article] (2023). Unlocking the enzymatic potential of lignocellulose depolymerization by wood-digesting marine shipworm Psiloteredo megatora.
• Votvik, Amanda Kristine; Kjendseth, Åsmund Røhr; Bissaro, Bastien; Stepnov, Anton; Sørlie, Morten & Eijsink, Vincent [Show all 7 contributors for this article] (2023). Structural and functional characterization of a cell-wall anchored bacterial lytic polysaccharide monooxygenase from Streptomyces coelicolor .
• Angeltveit, Camilla Fløien; Kommedal, Eirik; Eijsink, Vincent & Horn, Svein Jarle (2023). Light promotes lignin-catalyzed formation of hydrogen peroxide and LPMO activity.
• Balzer Le, Simone; Nguyen, Giang-Son; Jønsson, Malene; Aune, Ingvild; Streit, Wolfgang & Michalak, Leszek [Show all 11 contributors for this article] (2022). Functional screening of (meta)genomic libraries to discover novel enzymes and bioactive molecules for industrial applications.
• Nguyen, Giang-Son; Balzer Le, Simone; Jønsson, Malene; Aune, Ingvild; Cordas, Cristina M. & Moura, José J. G. [Show all 17 contributors for this article] (2022). Bioinformatics at SINTEF Biotechnology.
• Christensen, Idd Andrea; Stepnov, Anton; Eijsink, Vincent; Courtade, Gaston & Aachmann, Finn Lillelund (2022). Following the fate of lytic polysaccharide monooxygenases (LPMOs) under oxidative conditions by NMR spectroscopy.
• Tuveng, Tina Rise; Østby, Heidi; Stepnov, Anton; Vaaje-Kolstad, Gustav; Forsberg, Zarah & Eijsink, Vincent (2022). Effects of copper saturation on bacterial LPMOs active on chitin and cellulose.
• Stepnov, Anton; Forsberg, Zarah; Kjendseth, Åsmund Røhr; Sørlie, Morten & Eijsink, Vincent (2022). The effects of reductants and free copper ions on catalytic efficiency and enzyme stability in LPMO reactions.
• Eijsink, Vincent (2022). OxyMod research gives new opportunities for recirculating plastics. digitallifenorway.org.
• Eijsink, Vincent (2022). From single enzymes to enzyme systems – OxyMod leads the way. digitallifenorway.org.
• Eijsink, Vincent (2022). OxyMod PhD student unravels the kinetics of oxidative biomass conversion. digitallifenorway.org.
• Eijsink, Vincent (2022). Structure, function and application of LPMOs – unique monocopper peroxygenases for biomass processing and beyond.
• Eijsink, Vincent (2022). Structure and function of LPMOs –monocopper peroxygenases for biomass processing and beyond.
• Madland, Eva; Forsberg, Zarah; Wang, Yong; Lindorff-Larsen, Kresten; Niebisch, Axel & Modregger, Jan [Show all 9 contributors for this article] (2022). Structural and functional variation of two chitin-binding domains from CjLPMO10A.
• Duarte, R.S; Cordas, Cristina M.; Nguyen, Giang-Son; Valério, G.N.; Jønsson, Malene & Söllner, Katharina [Show all 9 contributors for this article] (2022). Characterization of a novel Dyp-type peroxidase from a marine actinobacterium.
• Cordas, Cristina M. (2022). A newly discovered Dyp-type peroxidase from a marine actinobacterium immobilization and electrochemical characterization.
• Cordas, Cristina M.; Valério, G.N.; Kjendseth, Åsmund Røhr; Eijsink, Vincent & Moura, José J.G. (2022). LPMO direct electrochemical characterization.
• Myskja, Bjørn Kåre & Myklebust, Alexander (2022). Missions, innovation and RR:Political framework for innovation-oriented research.
• Nguyen, Giang-Son; Balzer Le, Simone; Jønsson, Malene; Aune, Ingvild; Cordas, Cristina M. & Moura, José J. G. [Show all 14 contributors for this article] (2022). (Meta)genome mining for novel enzyme discovery.
• Nguyen, Giang-Son; Jønsson, Malene; Cordas, Cristina M.; Moura, José J. G.; Danso, Dominik & Streit, Wolfgang [Show all 7 contributors for this article] (2021). Mining of novel enzymes for biomass and plastic degradation in the Actinobacteria strain collection isolated from Trondheim's fjord, Norway.
• Myskja, Bjørn Kåre & Myklebust, Alexander (2021). Socratic dialogue on responsible innovation – a methodological experiment.
• Gírio, Francisco; Carvalheiro, Florbela & Eijsink, Vincent (2021). Enertech – Energy Technologies Fair .
• Martins, M.M; Carvalheiro, Florbela; Gírio, Francisco & Eijsink, Vincent (2021). Ethanol organosolv delignification of spruce biomass for biorefinery applications.
• Eijsink, Vincent (2021). Lytic Polysaccharide MonoOxygenases (LPMOs) - how chitin helped to change our views on enzymatic biomass conversion.
• Eijsink, Vincent (2021). Lytic Polysaccharide Monooxygenases (LPMOs) and other redox enzymes in polysaccharide degradation – fundamental and applied aspects.
• Martins, M.M; Carvalheiro, F & Gírio, F.M (2021). Ethanol organosolv delignification of spruce biomass for biorefinery applications.
• Courtade, Gaston; Forsberg, Zarah; Eijsink, Vincent & Aachmann, Finn Lillelund (2020). STRUCTURAL AND FUNCTIONAL INSIGHTS INTO THE MODE OF ACTION OF MODULAR LYTIC POLYSACCHARIDE MONOOXYGENASES.
• Eijsink, Vincent (2020). Biomass processing: Mechanistic aspects of lytic polysaccharide monooxygenases.
• Eijsink, Vincent (2020). Announcement of four recent publiucations in OxyMod.
• Wentzel, Alexander; Nguyen, Giang-Son & Degnes, Kristin Fløgstad (2020). Sequence- and function-based bioprospecting of Norwegian microbial biodiversity for novel natural products.
• Courtade, Gaston; Forsberg, Zarah; Wang, Yong; Pedersen, Martin N.; Buchinger, Edith & Heggset, Ellinor Bævre [Show all 10 contributors for this article] (2019). Structural and functional insights into the mode of action of a modular lytic polysaccharide monooxygenase.
• Eijsink, Vincent (2019). Introduction to the project.
• Eijsink, Vincent (2019). LPMOs - from chitin-binding proteins and sloppy endoglucanases to versatile (per)oxygenases for biomass processing.
• Kjendseth, Åsmund Røhr (2019). Mechanism of the chitinolytic peroxygenase reaction.
• Eijsink, Vincent (2019). Oxidative monocopper enzymes that accelerate glycoside hydrolase conversion of lignocellulosic biomass.
• Eijsink, Vincent (2019). Lytic polysaccharide monooxygenases (LPMOs) - unique mono-copper oxidases for biomass processing and more.
• Eijsink, Vincent (2019). Hydrogen Peroxide-Driven Oxidative Cleavage of Polysaccharides by Monocopper Enzymes.
• Courtade, Gaston; Forsberg, Zarah; Eijsink, Vincent & Aachmann, Finn Lillelund (2019). Structure and dynamics of a multi-domain lytic polysaccharide monosaccharide.
• Christensen, Idd Andrea; Courtade, Gaston; Le, Simone Balzer; Sætrom, Gerd Inger; Brautaset, Trygve & Aachmann, Finn Lillelund (2019). A New Expression System for Isotope Labelling of Proteins.
• Børve, Marta; Courtade, Gaston; Gaardløs, Margrethe & Aachmann, Finn Lillelund (2019). NMR study of enzyme kinetics for mannuronan C-5 epimerase AlgE4.
• Nguyen, Giang-Son; Jønsson, Malene; Neeraas, Priscilla C.; Eijsink, Vincent & Wentzel, Alexander (2019). Discovery and characterization of novel redox enzymes from Trondheim fjord's actinobacteria strain collection for lignocellulosic biomass degradation. Show summary

  Introduction Understanding enzymes and enzyme systems involved in lignocellulosic biomass degradation is the central topic of the OXYMOD project, funded by the Center of Digital Life Norway. Beside enzymes that are active against cellulose, redox enzymes such as laccases and peroxidases contribute to the process of breaking down lignin fraction. From long term bioprospecting activities by SINTEF and NTNU, a unique collection of actinobacteria strains has been established and 1200 strains have been sequenced using Illumina shotgun sequencing. The actinobacteria phylum has been known as a rich source of enzymes for lignin degradation and therefore is an attractive target for in silico mining of novel redox biocatalysts. Methods A customized workflow of data mining based on profiles HMM, multiple sequence alignment, and sequence similarity network has been developed and used for mining of new laccases and peroxidases from the database of 1200 actinobacteria genomes. The detected hits were further manually curated to have a shortlist of sequence candidates which were subsequently sent for gene synthesis and subcloned in expression vectors. The candidates were expressed, purified and the activity of the enzymes were tested against model substrate, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS). The laccases and peroxidases with confirmed ABTS activity were subsequently screened against a panel of small phenolic compounds as well as subjected to physiochemical characterization (pH optimum and stability, temperature stability). Result and discussion For mining of the new laccases, a custom profile HMM was created based on the sequences of two domain small laccases, SLAC-like family (Hfam39), obtained from Laccase and Multicopper Oxidase Engineering Database (LCCED). Four out of seven new laccase candidates were expressed in the soluble form and showed activity against ABTS after purification. Two enzymes, namely P06-A08 and P20-F12 appeared to be thermostable (activity was maintained even after two hours incubated at 80 °C). The pH optimum of the enzymes was identified in the range of pH 3.8 and 4.2 with the sharp decline of activity from pH 5 and forward. Among compounds tested for substrate scope study, activities against ferulic acid, sinapic acid, catechol, and 3,4-dihydroxybenzoic acid were confirmed. Most of identified peroxidase hits from the strain collection belong to the Dyp-type family (dye de colourising peroxidase). Like the new laccases, four peroxidases were active against ABTS after purification and have a pH optimum in the limited range of 3.0 and 3.4. The de colourising activity was confirmed using commercial dyes, Remazol Brilliant Blue R and Reactive Blue 5.

• Courtade, Gaston; Forsberg, Zarah; Vaaje-Kolstad, Gustav; Eijsink, Vincent & Aachmann, Finn Lillelund (2018). Structure and dynamics of a multi-domain lytic polysaccharide monooxygenase.
• Courtade, Gaston; Forsberg, Zarah; Heggset, Ellinor Bævre; Eijsink, Vincent & Aachmann, Finn Lillelund (2018). Structure and dynamics of a multi-domain lytic polysaccharide monosaccharide.
• Eijsink, Vincent (2018). Examples of enzyme technology and biorefining at NMBU.
• Eijsink, Vincent (2018). Bacteria – the new superheroes. Titan.uio.no.
• Eijsink, Vincent (2018). Bacteria – the new superheroes. digitallifenorway.org.
• Courtade, Gaston; Forsberg, Zarah; Heggset, Ellinor Bævre; Eijsink, Vincent & Aachmann, Finn Lillelund (2018). Cellulose oxidation by a modular LPMO.
• Courtade, Gaston; Forsberg, Zarah; Petrovic, Dejan; Sætrom, Gerd Inger; Heggset, Ellinor Bævre & Eijsink, Vincent [Show all 7 contributors for this article] (2018). NMR perspectives on Lytic Polysaccharide Monooxygenase.
• Bissaro, Bastien; Streit, Bennett; Isaksen, Ingvild; Beckham, Gregg T.; Eijsink, Vincent & DuBois, Jennifer (2018). Unravelling the molecular basis of the lytic polysaccharide (per)oxygenase reaction.
• Eijsink, Vincent & Vaaje-Kolstad, Gustav (2018). The 2018 Nobel Prize in Chemistry. NBS-nytt. ISSN 0801-3535. p. 34–35.
• Eijsink, Vincent; Sørlie, Morten; Kjendseth, Åsmund Røhr; Forsberg, Zarah & Stepnov, Anton (2022). An investigation of the interplay between in situhydrogen peroxide production and catalytic efficiency in lytic polysaccharide monooxygenase reactions. NMBU/KBM. ISSN 978-82-575-1900-1.
• Eijsink, Vincent; Østby, Heidi & Hennum, Karen (2022). Characterization of a Lytic Polysaccharide Monooxygenase from Schizophyllum commune belonging to Auxiliary Activity Family 9. NMBU/KBM.
• Myklebust, Alexander; Myskja, Bjørn Kåre; Carson, Siri Granum & Swierstra, Tsjalling (2022). A philosophical interpretation of the development of neo-Schumpeterian theory and policy. Norwegian University of Science and Technology. ISSN 2703-8084. 2022(132).
• Demmene, Camilla & Eijsink, Vincent (2020). Searching for a redox partner for bacterial lytic polysaccharide monooxygenases. NMBU.
• Marinovic, Adrian & Malmo, Jostein (2020). Establishment and validation of the XylS/Pm regulator/promoter system in Streptomyces lividans (delta)matAB for expression and secretion of Lytic Polysaccharide Monooxygenase. NTNU, Vectron Biosolutions.
• Malmo, Jostein & Horten, Kirsten (2020). Production of LPMO in Streptomyces. NTNU, Vectron Biosolutions.
• Votvik, Amanda Kristine & Eijsink, Vincent (2019). Studying substrate specificity by site-directed mutagenesis of a novel lytic polysaccharide monooxygenase. NMBU.
• Neeraas, Priscilla & Vaaje-Kolstad, Gustav (2019). Discovery, expression, and characterization of novel laccases derived from marine bacteria . NMBU.
• Siddqui, Sohaib & Malmo, Jostein (2019). Establishing Streptomyces lividans as a host for CBP21 expression: A proof of concept. NTNU, Vectron Biosolutions.
• Skogekker, Camilla & Malmo, Jostein (2018). Fundamental studies for the establishment of a XylS/Pm expression system in Streptomyces lividans. NTNU, Vectron Biosolutions.

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