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BIOTEK2021-Bioteknologi for verdiskaping

DL: OXYMOD - Optimized oxidative enzyme systems for efficient conversion of lignocellulose to valuable products

Alternative title: OXYMOD - Optimaliserte redoks-enzymsystemer for effektiv bearbeiding av lignocellulose

Awarded: NOK 20.0 mill.

In the OXYMOD project scientists with different competences work on discovering, understanding and further developing enzyme systems for the conversion of lignocellulose from Norwegian biomass into valuable products like sugars and aromatic chemicals. Redox enzymes have huge potential in the depolymerisation of cellulose, hemicellulose and lignin, but studying these enzymes is difficult and requires a systems approach. Thus, OXYMOD goes further than basic enzyme discovery, as we study the interplay between redox enzymes, their co-factors and their reaction partners, as well as the interplay between these novel enzymes and hydrolytic enzymes. OXYMOD studies these enzyme systems by looking at approx. 1000 marine Actinobacteria isolates whose genomes have been sequenced. OXYMOD combines life sciences (enzyme biochemistry, enzyme production technology, microbial biotechnology, high throughput screening, advanced analytics), ICT (bioinformatics, big data handling), mathematical sciences (enzyme systems and process modelling) and engineering (enzyme evolution, synthetic biology) for producing new biocatalytic systems for industrial application. Our focus is on developing enzyme solutions for the agricultural and forest sectors. Besides the enzymes and enzyme systems themselves, innovations from OXYMOD concern the generation of well-defined products streams, primarily sugars from (hemi-)cellulose and aromatic building blocks from lignin, for downstream applications (e.g. biofuels, bioplastics, feed). Additional applications can be envisaged, such as the development of green (= enzymatic) methods for producing fine chemicals. We have carried out large bioinformatic analyses to find enzyme candidates, which have been produced and characterized. A lot of energy was put into getting the enzymes produced, which is done by e.g. the company Vectron, who has managed to develop a new system for protein production in Streptomyces that is unique to Norway and very useful for several projects. We work with enzymes that in themselves have interesting applications and here we have made great progress (two PNAS papers). Based on these results, we have studied complete enzyme systems, i.e., mixtures of interacting enzymes, as they exist in nature. An unexpected amount of energy had to be used to study so-called abiotic factors (e.g., the presence of metal ions, redox active small molecules, and oxygen), because the project has shown that these factors have major effects on the efficiency of the enzyme systems. In order to understand and model our enzyme systems, the properties of the enzymes must be analyzed very carefully; therefore, we carry out a number of advanced studies of structure, dynamics and function (kinetics), partly in collaboration with our Portuguese project partners. We have a close dialogue with several interested companies and innovation experts from both Digital Life and NMBU. Both participate regularly in the project meetings. The project has an active RRI component, and a PhD student has submitted his dissertation that is based, in part, on RRI work in the project. Important progress in 2022 includes: - Publication of several articles and defense of a PhD thesis showing how redox enzyme systems can best be used in the bioprocessing of biomass. With NTNU, NMBU and SINTEF. - Good collaboration with our Portuguese partners that has led to several papers, including a fundamental paper that for the first time describes in-depth "cyclic voltammetry" of LPMOs, and a ground-breaking paper, appearing in Nature Communications, that shows that light also plays a role in these enzyme systems. Great joint publications. With NTNU, NMBU and SINTEF. - Massive new insights, partly NMR-based into how best to use lytic polysaccharide monooxygenases, one of the “hottest” current industrial enzymes. NTNU & NMBU. - Publication of an article on RRI-methodology. NTNU. - Successful defense of the RRI-related PhD. NTNU. - Progress in enzyme expression in Streptomyces, developed at Vectron. Highlight: successful expression and secretion of an LPMO and inducible expression of eGFP. With NTNU and SINTEF. - Based on insight from OxyMod, a large pilot trial is planned in St1's facilities where the findings from OxyMod will be tested in industrial scale. With NMBU. - Work is being done to follow up several other discoveries from OxyMod, including using lignin-modifying enzymes, which were discovered through "high-throughput screening" in OxyMod, in new bioprocesses.

Prosjektet har ført til nye nasjonale og internasjonale samarbeidskonstellasjoner som har ført til ny kunnskap og fellespublikasjoner. Vi har fått gjort ting, og gjort vitenskapelige framskritt som ville ha vært umulig uten dette prosjektet. Samspillet med Portugisiske partere har tilført en ny dimensjon tik norsk enzymforskning. Prosjektet har vært med på å danne grunnlaget for SFI Industriell Bioteknologi (2020-2028) men har også bidratt til å skape grunnlaget for andre prosjekter, deriblant et meget innovativt og tverrfaglig «blue-sky» ERC-Synergy prosjekt, hvor Eijsink (NMBU) er av fire PI (da andre PI er fra UiO, Italia og Tyskland). En annen follow-up er det nye Enzyclic prosjektet (ledet av Gustav Vaaje-Kolstad) hvor vi skal videreutvikle de redoksenzymene som vi har studert i OxyMod for bruk i enzymatisk resirkulering av plast. Prosjektet har gitt mye detaljert innsikt i hvordan redoksenzymer best kan anvendes i enzymatisk prosessering av platebiomasse og spesielt cellulose. Det har vært fortløpende dialoger med Borregaard og St1. Borregaard fullførte i 2020 et stort pilotforsøk (4000 liter skala) basert på kunnskap fra flere prosjekter, deriblant OxyMod. I 2022 har det vært en del samarbeid med St1 og pilotskala forsøk kan bli aktuelle i 2023. RRI arbeidet i prosjektet har ført til mye større bevissthet rundt, og kunnskap om, de prosessene som må til for å oppnå kunnskapsbasert innovasjon. Videre vil RRI arbeidet i OxyMod påvirke hvordan vi nærmer oss tverrfaglig forskning innen etikk, og gjøre oss i stand til å involvere prosjektpartnere mer direkte i empirisk etikkbasert forskning. Oxymod har videre ledet til et DLN/RoadMap to Innovation finansiert prosjekt som heter «InnoFree», med NMBU og Ard Innovation. Den vitenskapelige outputen fra OxyMod rundt enzymatisk prosessering av lignocellulose blir lagd merke til verden over og har vært presentert nasjonalt of internasjonalt gjennom flere plattformer.

The OXYMOD project will through a transdisciplinary approach define, develop and demonstrate applicability of new enzyme systems for the efficient biocatalytic conversion of lignocellulose from abundant Norwegian biomass into valuable products like sugars and aromatic building blocks. OXYMOD will focus on the still largely underexplored group of redox enzymes and their potential in the depolymerisation of cellulose, hemicellulose and lignin, including aspects such as redox enzyme interplay, co-factors and reaction partners, as well as their interplay with hydrolytic enzymes. OXYMOD will address these enzymes and enzyme systems as they occur and function in, among others, a unique in-house collection of approx. 1000 marine Actinobacteria isolates with genomes recently sequenced. Redox enzymes require co-factors and redox partners, and there is a considerable degree of cooperativity between different enzyme classes. Enzyme systems-scale understanding and eventually engineering the efficient degradation of lignocellulose by these enzyme systems, requires an integrated transdisciplinary approach far beyond 'simple' enzyme discovery. OXYMOD combines life sciences (enzyme biochemistry, enzyme production technology, microbial biotechnology, high throughput screening, advanced analytics), ICT (bioinformatics, big data handling), mathematical sciences (enzyme systems modelling, process modelling) and engineering (enzyme evolution, synthetic biology) for producing new and optimized biocatalytic systems for industrial application, primarily within the agricultural and forest sectors. Besides the enzymes and enzyme systems themselves, additional innovations from OXYMOD concern the generation of well-defined products streams, primarily sugars from (hemi-)cellulose and aromatic building blocks from lignin for a variety of downstream applications (e.g. biofuels & bioplastics).

Publications from Cristin

Funding scheme:

BIOTEK2021-Bioteknologi for verdiskaping

Thematic Areas and Topics

NordområdeneNordområdeneKlima, miljø og biologiske ressurserMarinBioteknologiMarin bioteknologiAvanserte produksjonsprosesserAvansert produksjonsteknologi som fag og teknologi (ny fra 2015)MarinMarin bioteknologiMiljøteknologiIKTIKT - Bruk og anvendelser i andre fagLTP3 Klima, polar og miljøBioteknologiLandbruksbioteknologiNaturmangfold og miljøMiljøteknologiLTP3 Hav og kystMiljøteknologiAnnen miljøteknologiPolitikk- og forvaltningsområderFiskeri og kystBioteknologiIndustriell bioteknologiLTP3 Marine bioressurser og havforvaltningNaturmangfold og miljøBioøkonomiInternasjonaliseringAvanserte produksjonsprosesserNaturmangfold og miljøSirkulær økonomiDelportefølje Et velfungerende forskningssystemBransjer og næringerProsess- og foredlingsindustriEtiske, juridiske og samfunnsmessige aspekterLTP3 IKT og digital transformasjonDelportefølje KvalitetLTP3 Et kunnskapsintensivt næringsliv i hele landetIKTIKT - Som fag og teknologiPolitikk- og forvaltningsområderSkog, landbruk og matKlimarelevant forskningLTP3 Fagmiljøer og talenterPortefølje Klima og miljøBransjer og næringerPolitikk- og forvaltningsområderLTP3 Høy kvalitet og tilgjengelighetDelportefølje InternasjonaliseringLTP3 Klima, miljø og energiPolitikk- og forvaltningsområderNæring og handelPortefølje ForskningssystemetIKTLTP3 Muliggjørende og industrielle teknologierPortefølje InnovasjonInternasjonaliseringInternasjonalt prosjektsamarbeidLTP3 Nano-, bioteknologi og teknologikonvergensPortefølje Muliggjørende teknologierBioteknologiPortefølje Banebrytende forskningGrunnforskningPortefølje Mat og bioressurserLTP3 Bioøkonomi og forvaltningLTP3 Styrket konkurransekraft og innovasjonsevneSirkulær økonomi