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 9780323992640. p. 163–189. doi: 10.1016/bs.mie.2022.08.049. 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 Show summary

  A considerable number of lytic polysaccharide monooxygenases (LPMOs) and other carbohydrate-active enzymes are modular, with catalytic domains being tethered to additional domains, such as carbohydrate-binding modules, by flexible linkers. While such linkers may affect the structure, function, and stability of the enzyme, their roles remain largely enigmatic, as do the reasons for natural variation in length and sequence. Here, we have explored linker functionality using the two-domain cellulose-active ScLPMO10C from Streptomyces coelicolor as a model system. In addition to investigating the WT enzyme, we engineered three linker variants to address the impact of both length and sequence and characterized these using small-angle X-ray scattering, NMR, molecular dynamics simulations, and functional assays. The resulting data revealed that, in the case of ScLPMO10C, linker length is the main determinant of linker conformation and enzyme performance. Both the WT and a serine-rich variant, which have the same linker length, demonstrated better performance compared with those with either a shorter linker or a longer linker. A highlight of our findings was the substantial thermostability observed in the serine-rich variant. Importantly, the linker affects thermal unfolding behavior and enzyme stability. In particular, unfolding studies show that the two domains unfold independently when mixed, whereas the full-length enzyme shows one cooperative unfolding transition, meaning that the impact of linkers in biomass-processing enzymes is more complex than mere structural tethering.

• 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. Full text in Research Archive 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.

• 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. doi: 10.1021/acssuschemeng.3c03714. Full text in Research Archive
• 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. 13(1), p. 5345–5345. doi: 10.1038/s41598-023-32263-7. 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. 13. doi: 10.1038/s41598-023-44278-1. Full text in Research Archive Show summary

  Lytic polysaccharide monooxygenases (LPMOs) are mono-copper enzymes that use O2 or H2O2 to oxidatively cleave glycosidic bonds. LPMOs are prevalent in nature, and the functional variation among these enzymes is a topic of great interest. We present the functional characterization of one of the 22 putative AA9-type LPMOs from the fungus Schizophyllum commune, ScLPMO9A. The enzyme, expressed in Escherichia coli, showed C4-oxidative cleavage of amorphous cellulose and soluble cello-oligosaccharides. Activity on xyloglucan, mixed-linkage β-glucan, and glucomannan was also observed, and product profiles differed compared to the well-studied C4-oxidizing NcLPMO9C from Neurospora crassa. While NcLPMO9C is also active on more crystalline forms of cellulose, ScLPMO9A is not. Differences between the two enzymes were also revealed by nuclear magnetic resonance (NMR) titration studies showing that, in contrast to NcLPMO9C, ScLPMO9A has higher affinity for linear substrates compared to branched substrates. Studies of H2O2-fueled degradation of amorphous cellulose showed that ScLPMO9A catalyzes a fast and specific peroxygenase reaction that is at least two orders of magnitude faster than the apparent monooxygenase reaction. Together, these results show that ScLPMO9A is an efficient LPMO with a broad substrate range, which, rather than acting on cellulose, has evolved to act on amorphous and soluble glucans.

• Myklebust, Alexander (2023). Innovasjon som kombinasjon, teknologi og omstilling. Norsk filosofisk tidsskrift. ISSN 0029-1943. 58(2-3). doi: 10.18261/nft.58.2-3.6. Full text in Research Archive Show summary

  Artikkelen drøftar korleis synet på teknologi og innovasjon har utvikla seg frå Joseph Schumpeter sin teori om økonomisk utvikling frå første halvdel av 1900-talet og fram til nyare tids innovasjonspolitikk som siktar mot å omstille samfunnet ved å peike ut samfunnsoppdrag. Ved å tolke utviklinga av innovasjonsomgrepet i lys av schumpeteriansk og ny-schumpeterianske teoriar om økonomisk utvikling, teknologisk innovasjon og rettferdig berekraftig omstilling undersøker artikkelen korleis innovasjonsomgrepet har utvikla seg saman med spørsmål om og korleis ein kan rasjonelt handtere dei sosiale og dei teknologiske endringane moderne samfunn stadig står overfor.

• 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 Show summary

  Background Cellulases play a key role in the enzymatic conversion of plant cell-wall polysaccharides into simple and economically relevant sugars. Thus, the discovery of novel cellulases from exotic biological niches is of great interest as they may present properties that are valuable in the biorefning of lignocellulosic biomass. Results We have characterized a glycoside hydrolase 5 (GH5) domain of a bi-catalytic GH5-GH6 multi-domain enzyme from the unusual gill endosymbiont Teredinibacter waterburyi of the wood-digesting shipworm Psiloteredo megotara. The catalytic GH5 domain, was cloned and recombinantly produced with or without a C-terminal family 10 carbohydrate-binding module (CBM). Both variants showed hydrolytic endo-activity on soluble substrates such as β-glucan, carboxymethylcellulose and konjac glucomannan, respectively. However, low activity was observed towards the crystalline form of cellulose. Interestingly, when co-incubated with a cellulose-active LPMO, a clear syn‑ ergy was observed that boosted the overall hydrolysis of crystalline cellulose. The crystal structure of the GH5 catalytic domain was solved to 1.0 Å resolution and revealed a substrate binding cleft extension containing a putative+3 subsite, which is uncommon in this enzyme family. The enzyme was active in a wide range of pH, temperatures and showed high tolerance for NaCl. Conclusions This study provides significant knowledge in the discovery of new enzymes from shipworm gill endo‑ symbionts and sheds new light on biochemical and structural characterization of cellulolytic cellulase. Study demon‑ strated a boost in the hydrolytic activity of cellulase on crystalline cellulose when co-incubated with cellulose-active LPMO. These findings will be relevant for the development of future enzyme cocktails that may be useful for the biotechnological conversion of lignocellulose.

• 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. 14(1). doi: 10.1038/s41467-023-36660-4. Full text in Research Archive Show summary

  Lytic polysaccharide monooxygenases (LPMOs) catalyze oxidative cleavage of crystalline polysaccharides such as cellulose and are crucial for the conversion of plant biomass in Nature and in industrial applications. Sunlight promotes microbial conversion of plant litter; this effect has been attributed to photochemical degradation of lignin, a major redox-active component of secondary plant cell walls that limits enzyme access to the cell wall carbohydrates. Here, we show that exposing lignin to visible light facilitates cellulose solubilization by promoting formation of H2O2 that fuels LPMO catalysis. Light-driven H2O2 formation is accompanied by oxidation of ring-conjugated olefins in the lignin, while LPMO-catalyzed oxidation of phenolic hydroxyls leads to the required priming reduction of the enzyme. The discovery that light-driven abiotic reactions in Nature can fuel H2O2-dependent redox enzymes involved in deconstructing lignocellulose may offer opportunities for bioprocessing and provides an enzymatic explanation for the known effect of visible light on biomass conversion.

• 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. 12(1). doi: 10.1038/s41598-022-10096-0. Full text in Research Archive
• 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. Full text in Research Archive
• 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. Full text in Research Archive
• 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 Show summary

  This article presents an experiment in using Socratic dialogue as a methodological approach to Responsible Research and Innovation (RRI) in an interdisciplinary life sciences research project. The approach seeks to avoid imposing a set of predetermined substantive norms by engaging the researchers in knowledge-seeking group discussions. We adapted Svend Brinkmann’s method of epistemic interviewing, in order to facilitate reflection on normative issues concerning responsibility in research and innovation in two research group sessions. Two elements characterize this approach, relating it to empirical ethics methodologies: (1) the aim is not to map and analyse opinions, but to develop knowledge based on the dialogue; and (2) the facilitators of the discussion are also active participants in the dialogue rather than mere “spectators”. Through a description of the approach and discussion of some key challenges, we show the method’s potential as a supplement to the catalogue of RRI approaches and argue that it serves a dual purpose of contributing to knowledge production and reflexivity.

• 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 Show summary

  Among the extensive repertoire of carbohydrate-active enzymes, lytic polysaccharide monooxygenases (LPMOs) have a key role in recalcitrant biomass degradation. LPMOs are copper-dependent enzymes that catalyze oxidative cleavage of glycosidic bonds in polysaccharides such as cellulose and chitin. Several LPMOs contain carbohydrate-binding modules (CBMs) that are known to promote LPMO efficiency. However, structural and functional properties of some CBMs remain unknown, and it is not clear why some LPMOs, like CjLPMO10A from the soil bacterium Cellvibrio japonicus, have multiple CBMs (CjCBM5 and CjCBM73). Here, we studied substrate binding by these two CBMs to shine light on their functional variation and determined the solution structures of both by NMR, which constitutes the first structure of a member of the CBM73 family. Chitin-binding experiments and molecular dynamics simulations showed that, while both CBMs bind crystalline chitin with Kd values in the micromolar range, CjCBM73 has higher affinity for chitin than CjCBM5. Furthermore, NMR titration experiments showed that CjCBM5 binds soluble chitohexaose, whereas no binding of CjCBM73 to this chitooligosaccharide was detected. These functional differences correlate with distinctly different arrangements of three conserved aromatic amino acids involved in substrate binding. In CjCBM5, these residues show a linear arrangement that seems compatible with the experimentally observed affinity for single chitin chains. On the other hand, the arrangement of these residues in CjCBM73 suggests a wider binding surface that may interact with several chitin chains. Taken together, these results provide insight into natural variation among related chitin-binding CBMs and the possible functional implications of such variation.

• 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. Full text in Research Archive
• Nguyen, Giang-Son; Lewin, Anna; Bartolomeo, Francesca di & 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 9783030583156. p. 439–471. doi: 10.1007/978-3-030-58315-6_14. Full text in Research Archive 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.

• 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. Full text in Research Archive 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.

• 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. Full text in Research Archive
• 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. Full text in Research Archive
• 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. 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.

• Bissaro, Bastien; Streit, Bennett; Isaksen, Ingvild; Eijsink, Vincent; Beckham, Gregg T. & DuBois, Jennifer [Show all 7 contributors for this article] (2020). Molecular mechanism of the chitinolytic peroxygenase reaction. Proceedings of the National Academy of Sciences of the United States of America (PNAS). ISSN 0027-8424. 117(3), p. 1504–1513. doi: 10.1073/pnas.1904889117. Full text in Research Archive
• 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 (PNAS). ISSN 0027-8424. 117(32), p. 19178–19189. doi: 10.1073/pnas.2004277117. Full text in Research Archive Show summary

  Lytic polysaccharide monooxygenases (LPMOs) have a unique ability to activate molecular oxygen for subsequent oxidative cleavage of glycosidic bonds. To provide insight into the mode of action of these industrially important enzymes, we have performed an integrated NMR/electron paramagnetic resonance (EPR) study into the detailed aspects of an AA10 LPMO–substrate interaction. Using NMR spectroscopy, we have elucidated the solution-phase structure of apo-BlLPMO10A from Bacillus licheniformis, along with solution-phase structural characterization of the Cu(I)-LPMO, showing that the presence of the metal has minimal effects on the overall protein structure. We have, moreover, used paramagnetic relaxation enhancement (PRE) to characterize Cu(II)-LPMO by NMR spectroscopy. In addition, a multifrequency continuous-wave (CW)-EPR and 15N-HYSCORE spectroscopy study on the uniformly isotope-labeled 63Cu(II)-bound 15N-BlLPMO10A along with its natural abundance isotopologue determined copper spin-Hamiltonian parameters for LPMOs to markedly improved accuracy. The data demonstrate that large changes in the Cu(II) spin-Hamiltonian parameters are induced upon binding of the substrate. These changes arise from a rearrangement of the copper coordination sphere from a five-coordinate distorted square pyramid to one which is four-coordinate near-square planar. There is also a small reduction in metal–ligand covalency and an attendant increase in the d(x2−y2) character/energy of the singly occupied molecular orbital (SOMO), which we propose from density functional theory (DFT) calculations predisposes the copper active site for the formation of a stable Cu–O2 intermediate. This switch in orbital character upon addition of chitin provides a basis for understanding the coupling of substrate binding with O2 activation in chitin-active AA10 LPMOs.

• 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. 23(12). doi: 10.1016/j.isci.2020.101785. Full text in Research Archive Show summary

  Heterologous expression of a biosynthesis gene cluster from Amycolatopsis sp. resulted in the discovery of two unique class IV lasso peptides, felipeptins A1 and A2. A mixture of felipeptins stimulated proliferation of cancer cells, while having no such effect on the normal cells. Detailed investigation revealed, that pre-treatment of cancer cells with a mixture of felipeptins resulted in downregulation of the tumor suppressor Rb, making the cancer cells to proliferate faster. Pre-treatment with felipeptins made cancer cells considerably more sensitive to the anticancer agent doxorubicin and re-sensitized doxorubicin-resistant cells to this drug. Structural characterization and binding experiments showed an interaction between felipeptins resulting in complex formation, which explains their synergistic effect. This discovery may open an alternative avenue in cancer treatment, helping to eliminate quiescent cells that often lead to cancer relapse.

• 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 Show summary

  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.

• 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. Full text in Research Archive 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.

• 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. p. 831–841. doi: 10.1021/acssuschemeng.9b04912. Full text in Research Archive
• 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. 12(235). doi: 10.1186/s13068-019-1578-5. Full text in Research Archive
• 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 Show summary

  Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that catalyze the oxidative cleavage of polysaccharides such as cellulose and chitin, a feature that makes them key tools in industrial biomass conversion processes. The catalytic domains of a considerable fraction of LPMOs and other carbohydrate-active enzymes (CAZymes) are tethered to carbohydrate-binding modules (CBMs) by flexible linkers. These linkers preclude X-ray crystallographic studies, and the functional implications of these modular assemblies remain partly unknown. Here, we used NMR spectroscopy to characterize structural and dynamic features of full-length modular ScLPMO10C from Streptomyces coelicolor. We observed that the linker is disordered and extended, creating distance between the CBM and the catalytic domain and allowing these domains to move independently of each other. Functional studies with cellulose nanofibrils revealed that most of the substrate-binding affinity of full-length ScLPMO10C resides in the CBM. Comparison of the catalytic performance of full-length ScLPMO10C and its isolated catalytic domain revealed that the CBM is beneficial for LPMO activity at lower substrate concentrations and promotes localized and repeated oxidation of the substrate. Taken together, these results provide a mechanistic basis for understanding the interplay between catalytic domains linked to CBMs in LPMOs and CAZymes in general.

• 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. Full text in Research Archive

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• Myklebust, Alexander (2025). The Meaning of Innovation. A Reflection on the Morality and Political Economy of Innovation. Springer Nature Switzerland. ISBN 9783032015518. doi: 10.1007/978-3-032-01551-8. Full text in Research Archive

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• Kommedal, Eirik; Angeltveit, Camilla Fløien; Sæther, Fredrikke; Klau, Leesa Jane; Aachmann, Finn Lillelund & Horn, Svein Jarle [Show all 7 contributors for this article] (2025). Natural photoredox catalysts promote light-driven LPMO activity and enzymatic turnover of biomass. Full text in Research Archive
• Varnai, Aniko & Eijsink, Vincent (2025). Recent insights into the activity, stability and diversity of LPMOs; ubiquitous enzymes performing amazing chemistry. Full text in Research Archive
• Varnai, Aniko (2025). LPMOs in biomass saccharification: What we can do already …and what we do not know yet how to do. Full text in Research Archive
• Forsberg, Zarah; Golten, Ole; Stepnov, Anton; Østby, Heidi; Tuveng, Tina Rise & Hall, Kelsi [Show all 10 contributors for this article] (2024). On the functional characterization of LPMOs; controlling reactions and understanding their outcome. Full text in Research Archive
• 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. Full text in Research Archive
• Angeltveit, Camilla Fløien; Kommedal, Eirik; Eijsink, Vincent & Horn, Svein Jarle (2023). Light promotes lignin-catalyzed formation of hydrogen peroxide and LPMO activity. Full text in Research Archive
• 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. Full text in Research Archive
• Redaksjon, KBM (2023). New knowledge about the role of light in the enzymatic breakdown of plant biomass published in Nature Communications. nmbu.no. Full text in Research Archive Show summary

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

• Nguyen, Giang-Son; Le, Simone Balzer; Aune, Ingvild Haugnes; Nærdal, Ingemar; Heggeset, Tonje Marita Bjerkan & Bartolomeo, Francesca Di [Show all 12 contributors for this article] (2023). Digital bioprospecting for exploring biodiversity and discovering novel bioactive compounds and industrial enzymes. Full text in Research Archive
• Nguyen, Giang-Son; Le, Simone Balzer; Aune, Ingvild Haugnes; Nærdal, Ingemar; Heggeset, Tonje Marita Bjerkan & Bartolomeo, Francesca Di [Show all 12 contributors for this article] (2023). Digital bioprospecting for exploring biodiversity and discovering novel bioactive compounds and industrial enzymes. Full text in Research Archive
• Eijsink, Vincent (2022). Structure and function of LPMOs –monocopper peroxygenases for biomass processing and beyond. Full text in Research Archive
• Eijsink, Vincent (2022). Structure, function and application of LPMOs – unique monocopper peroxygenases for biomass processing and beyond. Full text in Research Archive
• Nguyen, Giang-Son; Le, Simone Balzer; 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. Full text in Research Archive
• Le, Simone Balzer; 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. Full text in Research Archive
• 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. Full text in Research Archive
• Eijsink, Vincent (2022). OxyMod research gives new opportunities for recirculating plastics. digitallifenorway.org. Full text in Research Archive
• Eijsink, Vincent (2022). From single enzymes to enzyme systems – OxyMod leads the way. digitallifenorway.org. Full text in Research Archive
• Eijsink, Vincent (2022). OxyMod PhD student unravels the kinetics of oxidative biomass conversion. digitallifenorway.org. Full text in Research Archive
• Myskja, Bjørn Kåre & Myklebust, Alexander (2022). Missions, innovation and RR:Political framework for innovation-oriented research. Full text in Research Archive
• Nguyen, Giang-Son; Le, Simone Balzer; Jønsson, Malene; Aune, Ingvild; Cordas, Cristina M. & Moura, José J. G. [Show all 17 contributors for this article] (2022). Bioinformatics at SINTEF Biotechnology. Full text in Research Archive
• 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. Full text in Research Archive
• 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. Full text in Research Archive
• 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. Full text in Research Archive
• 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. Full text in Research Archive
• Cordas, Cristina M. (2022). A newly discovered Dyp-type peroxidase from a marine actinobacterium immobilization and electrochemical characterization. Full text in Research Archive
• Cordas, Cristina M.; Valério, G.N.; Kjendseth, Åsmund Røhr; Eijsink, Vincent & Moura, José J.G. (2022). LPMO direct electrochemical characterization. Full text in Research Archive
• 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. Full text in Research Archive
• Eijsink, Vincent (2021). Lytic Polysaccharide MonoOxygenases (LPMOs) - how chitin helped to change our views on enzymatic biomass conversion. Full text in Research Archive
• Martins, M.M; Carvalheiro, Florbela; Gírio, Francisco & Eijsink, Vincent (2021). Ethanol organosolv delignification of spruce biomass for biorefinery applications. Full text in Research Archive
• Martins, M.M; Carvalheiro, F & Gírio, F.M (2021). Ethanol organosolv delignification of spruce biomass for biorefinery applications. Full text in Research Archive
• Eijsink, Vincent (2021). Lytic Polysaccharide Monooxygenases (LPMOs) and other redox enzymes in polysaccharide degradation – fundamental and applied aspects. Full text in Research Archive
• Myskja, Bjørn Kåre & Myklebust, Alexander (2021). Socratic dialogue on responsible innovation – a methodological experiment. Full text in Research Archive
• Courtade, Gaston; Forsberg, Zarah; Eijsink, Vincent & Aachmann, Finn Lillelund (2020). STRUCTURAL AND FUNCTIONAL INSIGHTS INTO THE MODE OF ACTION OF MODULAR LYTIC POLYSACCHARIDE MONOOXYGENASES. Full text in Research Archive
• Wentzel, Alexander; Nguyen, Giang-Son & Degnes, Kristin Fløgstad (2020). Sequence- and function-based bioprospecting of Norwegian microbial biodiversity for novel natural products. Full text in Research Archive
• Eijsink, Vincent (2019). Lytic polysaccharide monooxygenases (LPMOs) - unique mono-copper oxidases for biomass processing and more. Full text in Research Archive
• Kjendseth, Åsmund Røhr (2019). Mechanism of the chitinolytic peroxygenase reaction. Full text in Research Archive
• Eijsink, Vincent (2019). Oxidative monocopper enzymes that accelerate glycoside hydrolase conversion of lignocellulosic biomass. Full text in Research Archive
• Eijsink, Vincent (2019). LPMOs - from chitin-binding proteins and sloppy endoglucanases to versatile (per)oxygenases for biomass processing. Full text in Research Archive
• Eijsink, Vincent (2019). Hydrogen Peroxide-Driven Oxidative Cleavage of Polysaccharides by Monocopper Enzymes. Full text in Research Archive
• Eijsink, Vincent (2019). Introduction to the project. Full text in Research Archive
• 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. Full text in Research Archive 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; Eijsink, Vincent & Aachmann, Finn Lillelund (2019). Structure and dynamics of a multi-domain lytic polysaccharide monosaccharide. Full text in Research Archive
• 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. Full text in Research Archive
• Børve, Marta; Courtade, Gaston; Gaardløs, Margrethe & Aachmann, Finn Lillelund (2019). NMR study of enzyme kinetics for mannuronan C-5 epimerase AlgE4. Full text in Research Archive
• 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. Full text in Research Archive
• Courtade, Gaston; Forsberg, Zarah; Heggset, Ellinor Bævre; Eijsink, Vincent & Aachmann, Finn Lillelund (2018). Structure and dynamics of a multi-domain lytic polysaccharide monosaccharide. Full text in Research Archive
• Eijsink, Vincent & Vaaje-Kolstad, Gustav (2018). The 2018 Nobel Prize in Chemistry. NBS-nytt. ISSN 0801-3535. p. 34–35. Full text in Research Archive
• Bissaro, Bastien; Streit, Bennett; Isaksen, Ingvild; Beckham, Gregg T.; Eijsink, Vincent & DuBois, Jennifer [Show all 7 contributors for this article] (2018). Unravelling the molecular basis of the lytic polysaccharide (per)oxygenase reaction. Full text in Research Archive
• 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. Full text in Research Archive
• Courtade, Gaston; Forsberg, Zarah; Heggset, Ellinor Bævre; Eijsink, Vincent & Aachmann, Finn Lillelund (2018). Cellulose oxidation by a modular LPMO. Full text in Research Archive
• Eijsink, Vincent (2018). Examples of enzyme technology and biorefining at NMBU. Full text in Research Archive
• Courtade, Gaston; Forsberg, Zarah; Vaaje-Kolstad, Gustav; Eijsink, Vincent & Aachmann, Finn Lillelund (2018). Structure and dynamics of a multi-domain lytic polysaccharide monooxygenase. Full text in Research Archive
• Eijsink, Vincent (2018). Bacteria – the new superheroes. [Journal]. Titan.uio.no. Full text in Research Archive
• Eijsink, Vincent (2018). Bacteria – the new superheroes. [Journal]. digitallifenorway.org. Full text in Research Archive
• Eijsink, Vincent; Sørlie, Morten; Kjendseth, Åsmund Røhr; Forsberg, Zarah & Stepnov, Anton (2022). An investigation of the interplay between in situ hydrogen peroxide production and catalytic efficiency in lytic polysaccharide monooxygenase reactions. Norges miljø- og biovitenskapelige universitet (NMBU). ISSN 9788257519001. Full text in Research Archive Show summary

  Lytisk polysakkarid monooksygenaser (LPMOer) er mono-kobberenzymer som kan katalysere oksidativ depolymerisering av krystallinske substrater som kitin og cellulose. LPMOer er av høy interesse for både forskere og industri på grunn av deres unike evne til å virke på den krystallinske overflaten til substratene fremfor på de isolerte polysakkaridkjedene. Selv om mye forskning er gjennomført, er det fremdeles mye usikkerhet forbundet med mekanismen og kinetikken av LPMOkatalyserte reaksjoner. Opprinnelig trodde man at LPMOer benytter molekylært oksygen som ko-substrat, men i senere tid er det blitt vist at enzymene foretrekker hydrogenperoksid og at de derfor fungerer som peroksygenaser istedenfor monooksygenaser. Peroksygenasereaksjonen vil trolig skje selv ved fravær av tilsatt H2O2 fordi de fleste aerobe reaksjonsbetingelsene vil føre til at det dannes H2O2 in situ, for eksempel som følge av LPMOenes oksidaseaktivitet. Denne studien har satt søkelys på det komplekse samspillet mellom H2O2-genererende reaksjoner og den katalytiske effektiviteten til LPMOer som bryter ned cellulose i reaksjoner med oksygen og et utvalg av reduktanter.

• Eijsink, Vincent; Østby, Heidi & Hennum, Karen (2022). Characterization of a Lytic Polysaccharide Monooxygenase from Schizophyllum commune belonging to Auxiliary Activity Family 9. Norges miljø- og biovitenskapelige universitet (NMBU). Full text in Research Archive Show summary

  Biomass is a promising feedstock to reduce our dependence on fossil reserves. By using biomass, such as lignocellulose, products can be produced more climate-friendly and are therefore of large industrial interest. Biomass is also renewable, in contrast to fossil reserves, like oil and coal. However, the downside of biomass is that lignocellulose and other polysaccharides can be a challenge to degrade because of their crystalized and insoluble structure. Previously, hydrolytic enzymes were thought to be the only enzymes degrading polysaccharides. However, in 2010, a new paradigm was born, when researchers first described a protein that degraded chitin through oxidative cleavage. Subsequently, the enzymes have been characterized as lytic polysaccharide monooxygenases (LPMOs), and they do not only work on chitin, but also on cellulose and other recalcitrant polysaccharides. The discovery of LPMOs was a big step in the direction toward more efficient degradation of recalcitrant polysaccharides. LPMOs act on parts of the substrate that hydrolytic enzymes cannot properly degrade, thus making recalcitrant polysaccharides more available for traditional hydrolases by creating free chain ends. LPMOs are oxidative mono-copper enzymes that can oxidatively cleave glycosidic bonds in polysaccharides. To complete a catalytic cycle, the active site copper Cu(II) must be reduced to Cu(I), and the LPMOs require a dioxygen co-substrate, either O2 or H2O2. Since LPMOs show variation in their ability to attack various forms of cellulose, it is important for the industry to have a wide spectrum of LPMOs to use on different substrates. Therefore, ScLPMO9A from Schizophyllum commune, which is one of several LPMOs that have not been completely characterized yet, was studied, and compared with NcLPMO9C from Neurospora crassa for this thesis. ScLPMO9A and NcLPMO9C are both exclusively C4 oxidizing LPMOs, acting on soluble substrates. Results from this study show, for example, that ScLPMO9A depolymerises various forms of cellulose and hemicelluloses and that the substrate specificity and product formation differ from NcLPMO9C. For example, ScLPMO9A is active on xyloglucan, glucomannan and soluble cello-oligosaccharides, but not active on more crystalline forms of cellulose, like Avicel, while NcLPMO9C is active on more crystalline forms of cellulose. In addition, they also show that the reaction rate is increased by adding H2O2 to the reactions for both ScLPMO9A and NcLPMO9C. The results from this thesis give important information about the biochemistry of ScLPMO9A, that can be used for further studies.

• Myklebust, Alexander; Myskja, Bjørn Kåre; Carson, Siri Granum & Swierstra, Tsjalling (2022). A philosophical interpretation of the development of neo-Schumpeterian theory and policy. Norges teknisk-naturvitenskapelige universitet. ISSN 9788232663408. Full text in Research Archive Show summary

  English summary This thesis is a philosophical interpretation of a set of concepts and ideas that are influencing contemporary innovation discourse and innovation policies. The thesis starts from German philosopher Jürgen Habermas’ argument that one of the roles that the philosopher can play in the modern world is the role of interpreter of expert knowledge. From this starting point, the thesis centres on the so-called neo-Schumpeterian strand of innovation studies. Following in the footsteps of Austrian economist and sociologist Joseph Schumpeter (1883–1950), neo-Schumpeterians have emphasised the cyclical nature of capitalist development and innovation patterns. By reading and analyzing texts by three central economic thinkers (Joseph Schumpeter, Christopher Freeman and Mariana Mazzucato), as well as two sets of innovation policy documents (the expert reports leading up to “Horizon Europe” and the Oslo Manual), the thesis aims to spell out theoretical and normative ideas that guide contemporary neo-Schumpeterian theory and policy. The thesis ends up with three key findings about how certain Schumpeterian ideas have been developed by Freeman and Mazzucato, and adapted to specific innovation policy contexts. The first key finding concerns the notion of creativity. Schumpeter attributed to the individual entrepreneur the ability creatively respond to economic downturns, by carrying out an innovation, and thereby setting a new direction for economic production and activity. The thesis argues that the Schumpeterian notions of creativity, albeit in altered forms, exist in Freeman’s notion of national systems of innovation and in Mazzucato’s notion of the entrepreneurial state. The second key finding concerns the concept of the entrepreneur. Schumpeter primarily saw entrepreneurs as individuals who challenge the existing ways of doing things. Freeman, however, went in the direction of attributing the entrepreneurial function to systems of innovation, while Mazzucato has argued that it is the state that holds this function. The third key finding concerns the nature of development in modern capitalist societies. While Schumpeter thought that development was generated from within the capitalist system itself, current innovation policies of transformative change assume that capitalism is transformed by concerted state government efforts.

• Demmene, Camilla & Eijsink, Vincent (2020). Searching for a redox partner for bacterial lytic polysaccharide monooxygenases. Norges miljø- og biovitenskapelige universitet (NMBU). Full text in Research Archive Show summary

  Biomass is considered a vast resource for supplying sustainable and renewable energy. Lignocellulose is the most abundant source of biomass found on earth and utilization of this energy source is studied around the world. Enzymatic degradation is a promising approach to produce fuels, chemicals and materials from lignocellulosic biomass. A wide array of biomass-degrading enzymes can be found in nature, e.g. in white- and brown rot fungi, or in bacteria such as Cellvibrio japonicus. The discovery of lytic polysaccharide monooxygenases (LPMOs) has been of tremendous importance in understanding and utilizing biomass degradation processes. LPMOs are oxidative mono-copper enzymes that degrade glycosidic bonds in recalcitrant polysaccharides such as cellulose and chitin. The LPMOs require an initial reduction of the copper active site and a dioxygen co-substrate (e.g. O2 or H2O2) before catalysis can occur. In vitro studies commonly use reducing agents such as ascorbic acid to facilitate this reduction. In fungi, several redox enzymes that can directly interact with and reduce the LPMO active site have been described. In addition, several of those redox partner enzymes can also produce and supply the LPMO with H2O2. However, for bacterial LPMOs no similar redox partners have been identified so far, although a few potential candidates have been suggested. This study examines the potential redox ability of two such candidates, hereinafter called LPMO activating protein A and B (LapA and LapB), from the marine bacterium Saccharophagus degradans. These proteins were tested for their potential ability to drive the LPMO from the same organism, called SdLPMO10A. In addition, this novel tri-modular LPMO was subjected to functional characterization where it was shown to be a C1-oxidizing, cellulose-active enzyme with optimal activity at 60 °C. The apparent melting temperature of the copper-loaded enzyme was measured to 57 °C and 47 °C when the copper was removed from the active site. The C-terminal carbohydrate-binding domain was shown to bind cellulose, but the internal domain is still of unknown function and calls for further characterization. LapA and LapB are also multi-modular enzymes which have similar sequence- and domain organization as two proteins that has been shown to be co-expressed with LPMOs and are important for optimal growth on cellulose in the bacterium Cellvibrio japonicus. Like SdLPMO10A, LapA bound to cellulose, but LapB did not show any binding which may be a result of incorrect folding during a denaturation/refolding process. Initial activity tests of LapA and LapB to drive SdLPMO10A did not result in desired oxidative LPMO activity during the tested conditions. There are many factors which have yet to be explored, the main being to investigate the occurrence of a potent redox cofactor present in LapA and LapB. For the time being, LapA and LapB remain potential candidates for bacterial redox partners of LPMOs.

• 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. Full text in Research Archive
• Malmo, Jostein & Horten, Kirsten (2020). Production of LPMO in Streptomyces. NTNU, Vectron Biosolutions. Full text in Research Archive
• Votvik, Amanda Kristine & Eijsink, Vincent (2019). Studying substrate specificity by site-directed mutagenesis of a novel lytic polysaccharide monooxygenase. Norges miljø- og biovitenskapelige universitet (NMBU). Full text in Research Archive Show summary

  Cellulose and chitin are carbon-rich biopolymers which constitute the two largest biomass resources on earth. Conversion of these non-edible and renewable resources into useful products of value is essential to secure a sustainable bioeconomy. However, the efficient processing of cellulose and chitin is hampered by their recalcitrant nature. To exploit the energy-rich composition of these homopolymers, different microorganisms have evolved specialized enzyme systems which include enzymes referred to as lytic polysaccharide monooxygenases (LPMOs). LPMOs are copper-dependent enzymes, which in the presence of an external electron donor, catalyzes cleavage of the glycosidic linkages in recalcitrant polysaccharides through an oxidative and somewhat enigmatic reaction mechanism. LPMOs display relatively flat substrate binding surfaces which enables them to bind to the planar surface of crystalline polysaccharides. Various structural motifs, of conserved surface exposed residues found on the substrate binding face, are believed to contribute the specific oxidizing activity of LPMOs toward different polysaccharides. Seven LPMO encoding genes are found within the genome of the Gram-negative soil bacterium Streptomyces coelicolor. Among these LPMOs, various specificities for substrate and regioselectivities have been identified such as C1- or C1/C4-spesific cellulose oxidizing enzymes, and enzymes which display activity both toward cellulose (C1/C4) and chitin (C1). One of these enzymes which have not been characterized yet, called ScLPMO10D, exhibits a catalytic domain with conserved motifs specific for both chitin- and cellulose oxidizing LPMOs. In addition, ScLPMO10D exhibit a C-terminal domain that have been predicted to covalently anchor the catalytic domain to the cell wall, an unusual feature which may classify ScLPMO10D within a novel clade of Auxiliary Activity Family 10 (AA10) enzymes. In this study, ScLPMO10D was characterized as a chitin oxidizing enzyme with particular specificity toward β-chitin. Taking advantage of its sequence similarity to cellulose-oxidizing AA10s, five surface-exposed residues were targeted for site-directed mutagenesis in an attempt to change the substrate specificity toward cellulose. Out of the six mutants produced, none displayed activity toward cellulose. However, out of two mutants with retained activity on β- chitin, one displayed an initial rate of catalytic activity which exceeded the wild type enzyme.

• Neeraas, Priscilla & Vaaje-Kolstad, Gustav (2019). Discovery, expression, and characterization of novel laccases derived from marine bacteria. Norges miljø- og biovitenskapelige universitet (NMBU). Full text in Research Archive Show summary

  Lignocellulose er den mest rikelige formen for biomasse på jorda og er primært bygget opp av tre organiske polymerer; cellulose, hemicellulose, og lignin. Veletablerte teknologier eksisterer for å produsere nyttige produkter slik som brensel og kjemikalier fra cellulose og hemicellulose. Lignin, derimot, har i dag begrenset bruk som nyttelses produkter. Hvert år produseres det store mengder lignin som industrielt biprodukt, spesielt i papir industrien hvor omtrent 98% av materialet blir brent for energiproduksjon. Avhengigheten av fossile ressurser for å lage brensel, kjemikalier, og forbruksvarer har ødeleggende effekt på miljøet, og økende bekymring har resultert i økt etterspørsel etter grønnere og mer bærekraftige produkter. I en bioraffineri setting har lignin potensiale til å bli brukt i produksjonen av en rekke bio-baserte kjemikalier og materialer, og bruk av enzymer for lignin foredling er sett på som en lovende tilnærming i bioraffineri industrien.

• Siddqui, Sohaib & Malmo, Jostein (2019). Establishing Streptomyces lividans as a host for CBP21 expression: A proof of concept. NTNU, Vectron Biosolutions. Full text in Research Archive
• Skogekker, Camilla & Malmo, Jostein (2018). Fundamental studies for the establishment of a XylS/Pm expression system in Streptomyces lividans. NTNU, Vectron Biosolutions. Full text in Research Archive

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