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Creating ad hoc enzymes for the production of the SUSBIND renewable bio-binders

Several thousand putative UPO (unspecific peroxygenases) sequences have been found in genetic databases and fungal genomes, indicating their widespread occurrence in the whole fungal kingdom, but only a handful of UPOs have been characterized so far, and almost nothing substantial is known on their natural function(s).

Their reliable heterologous expression remains tricky, despite all recent progress in this field.

Caption: Sections of wild peroxygenase (left) and a mutated variant (right) displaying the differences in their substrate access channels leading to the heme cofactor (red sticks). The engineered variant bears two phenylalanines (highlighted in aquamarine) that constrain the channel, switching the regioselectivity of the enzyme towards selective monoepoxide production from polyunsaturated fatty acids.

As a result of international collaboration within SUSBIND project, the CSIC-CIB-IRNAS (Spain), the Barcelona Supercomputing Center, the Technical University of Dresden (Germany) and the SME JenaBios (Germany) recently published in ACS Catalysis the development of novel fungal peroxygenases, which behave as “P450s with advantages”.

Both P450s and peroxygenases are enzymes that catalyse the oxygenation — the introduction of oxygen atoms — of a variety of substrates. Such reactions are very difficult to perform by chemical means, especially when the substrate is aliphatic, as is the case of fatty acids. Moreover, the use of enzymes is generally much more environmentally friendly than that of chemical catalysts. Enzymes are active under mild conditions, have a natural origin and are normally very selective in the catalysis, giving rise to fewer undesired by-products.

So, what is the meaning of peroxygenases which behave as “P450s with advantage”? Which is the advantage? Is the behaviour as a biocatalyst for oxygenation processes as a result of their self-sufficient monooxygenase activity? Yes, unspecific peroxygenases rely on H2O2 for catalysis, while P450s normally need other enzymatic partners and are less stable due to their intracellular nature. Furthermore, the study results give deeper insight into describing the structural basis for the catalytic properties on different fatty acids and, besides, show the potential to produce compounds of high-added value at preparative scale. As Municoy M. et al. have used powerful computational tools, such as the software PELE for ligand diffusion into proteins to simulate the behaviour of the enzymes on their substrates, being able to satisfactorily predict the oxygenation type (epoxidation versus hydroxylation) of the new peroxygenases. Computational data have been combined with the experimental analysis of the oxygenated products formed and process scale-up. Factors related to the enzymatic active-site architecture and the double-bond distribution in the different fatty acids were suggested by the computational analyses identified in experimental reactions and confirmed by site-directed mutagenesis of the heme access channel. In this way, the structural determinants for the catalytic properties of the different enzymes analysed have been unveiled.

The potential of this combined computational-experimental approach opens the door for new biotechnological processes for the enzymatic production of bio-based chemicals, which was further demonstrated by the preparative regio- and stereoselective epoxidation of α-linolenic acid. The product of this reaction, cis,cis-15(R),16(S)-octadeca-9,12-dienoic acid, was obtained by employing wild and engineered peroxygenases attaining 80−83% enantiomeric excess and over 99% regioselectivity. This, and other monoepoxides, from polyunsaturated fatty acids are nearly impossible to be obtained by other chemical means.

Therefore, their enzymatic synthesis developed as a final output of the study, represents a new and interesting reaction given the biological activity of these compounds, in addition to their application in organic chemistry as highly reactive molecules.

Confirming that epoxides of unsaturated fatty acids produced using the above-mentioned enzymes and vegetable oil feedstocks is a valuable finding within SUSBIND in its aim to develop a bio-based adhesive.

Reference: Fatty-acid oxygenation by fungal peroxygenases: From computational simulations to preparative regio- and stereoselective epoxidation. Municoy M., González-Benjumea A., Carro J., Aranda C., Linde D., Renau-Mínguez C., Ullrich R., Hofrichter M., Guallar V., Gutiérrez A., and Martínez A.T. 2020. ACS Catal.  http://dx.doi.org/10.1021/acscatal.0c03165

 

“Biotechnology a Driver for Clean & Healthy Circular Economies – The Most Innovative EU Biotech-Projects”

SUSBIND – Nomination to be one of the most innovative biotech-projects in the EU by the KETBIO project platform!

“KETBIO’s  experts and its Commercial Committee have short-listed the top 10 biotechnology projects out of more than 300 EU-funded research projects. Please cast your vote online to decide on the most impactful and most promising EU-funded research in key enabling biotechnologies – from alternative proteins, bio-based compounds and food packs to microbial plastic recycling, enzymatic tools and industrial biotechnology.”

(C) KETBIO “Biotech Innovation at the Heart of a Green and Healthy Recovery for Europe” / Project Nominations

 

The list of projects comprises:

  • Application of cold plasma treatment for antimicrobial contact lenses
  • A new generation of microbial electrochemical wetland for effective decentralized wastewater treatment
  • Exploiting native endowments by re-factoring, re-programming and implementing novel control loops in Pseudomonas putida for bespoke biocatalysis
  • Fully bio-based and bio-degradable ready meal packaging
  • From plastic waste to plastic value using Pseudomonas putida synthetic biology
  • Re-think all plastic packaging – Wood-based fructose for production of plastic bottles and all plastic packaging
  • Sustainable jet fuel from flexible waste biomass
  • Industrial applications of marine enzymes: Innovative screening and expression platforms to discover and use the functional protein diversity from the sea
  • Innovative and scalable biotechnology using microbial fuel cell and anaerobic digestion for the treatment of micro-scale industrial and agriculture effluents to recover energy from waste

Many thanks again for your vote & help to make an impact for EU’s key enabling biotechnologies!

 

KETBIO Online Booster Conference Participation

SUSBIND, nominated for the Top-10 Most Innovative Biotech Projects in the EU, has been invited to give a talk at the KETBIO “Biotech Innovation at the Heart of a Green and Healthy Recovery for Europe” Online Booster Conference, June 17th. Among several invited speakers from the European Commission & Innovation Council, industry, policy makers, investors and commercial experts, RTDS represented SUSBIND in the afternoon session, accompanied by two other finalist projects.

“Biotech paves the way towards a broad range of applications including new sustainable consumer products such as bio-plastics, bio-based paints and glues, detergents, fertilizers for agriculture and sustainable fuels and energy carriers”, said Juergen Tiedje (European Commission, Head of Unit Sustainable Industry Systems), elaborating on KETBIO’s project analysis as baseline for the commercialisation and innovation potential.

The KETBIO project prior to the release of the Green Deal, tackling today’s challenges with knowledge by gathering best ideas and sustainable technologies for the modernisation of industrial sectors to become more circular. Industrial biotechnology has received and is still receiving significant funding from the EU, as it is seen as a key enabling technology for many fields, however time-to-market is a lengthy process and a lot of research and start-ups are disappearing into the “valley of death”, observes Tiedje, emphasizing the role of funding and additional services to support innovative projects.

“Overall, economy is mainly linear, and only a very little part is circular, which is unsustainable in a long-term view. The solution is: bioeconomy – bio-based materials replace fossil-based resources with less emissions and waste. It is intrinsically more efficient than the linear economy”, added Pavel Misiga (European Commission, Head of the Circular Economy and Bio-based Systems Unit, Directorate “Healthy Planet”) and on the EU support to deployment of bio-based solutions and transition in the next periods: “It can be expected that there will be a growth of the bio-based economy as a consequence of the demand for bio-based solutions. A part of the support is provided by the Bio-Based Industry Joint Undertaking public-private partnership for example. The circular bio-based Europe programme under Horizon Europe with the InvestEU financial instrument provides €11 billion for research & innovation investments.”

In total, the InvestEU is expected to mobilise at least €650 billion in additional investment between 2021 and 2027, to support four main policy areas: sustainable infrastructure; research, innovation and digitisation; small and medium businesses; and social investment and skills.

 

KETBIO: “A novel cluster model to bring KEY ENABLING BIOTECHNOLOGY research closer to markets and society”

The KETBIO project hub has attracted 1200 users over the last three years, says coordinator Dr. Kathrin Rübberdt (DECHEMA Gesellschaft für chemische Technik und Biotechnologie E.V.). In fact, KETBIO is an EU project itself, receiving funding from the European Union’s Horizon 2020 Research & Innovation Programme. Since October 2017, KETBIO supported Europe’s bioeconomy projects by providing an online platform for networking and knowledge exchange between research-industry-policy stakeholders, as well as organising conferences and webinars.

“This KETBIO project aims at establishing a novel cluster model of biotechnology research projects under HORIZON 2020 to enhance and demonstrate the impact and the outreach of EU funded key enabling biotechnology research. All impact of research is reflected in an appropriate up-take of research outputs through business and society at large as well as through integration into technological and societal systems, the here proposed novel clustering model will thus act as a pivot trajectory to achieve a maximum of these goals for the biotechnology: The proposed cluster will strive to further Research & Technical Development and innovation through networking and alliance forming and through capacity gains of cluster members. The envisaged clustering of projects and linkage to knowledge transfer activities will allow accelerated industrial exploitation of results through partnering and will maximise impact through exploiting synergies in knowledge transfer and communication. Supporting and coordination activities of KETBIO will lead to the set-up of an actively managed cluster-network of projects facilitating sharing of insights, mutual learning, working group exchange, partnering with industry, dissemination of results and exploring of exploitation pathways.” (CORDIS)

 

Last but not least, we are delighted to announce that SUSBIND is part of the KETBIO Flagship BookletA project showcase that is available online now!

(C) KETBIO Flagship Projects (online booklet)

 

Author: Dr. Stefan Weiss

Dissemination & Communication / RTDS Group

 

Further events with SUSBIND participation:

Stakeholderdialog Biobased Industry (Vienna, Austria)

BBI JU Stakeholder Forum 2019 (Brussels, Belgium)

European Summit of Industrial Biotechnology 2019 (Graz, Austria)

European Summit of Industrial Biotechnology 2019

The Austrian Centre of Industrial Biotechnology (acib GmbH) organized the European Summit of Industrial Biotechnology” (esib) for the fourth time, gathering the European biotechnology research and industry community on this communication platform for industrial biotechnology. The event not only covers science, but also addresses industrial needs and economic demands, funding resources and political aspirations, and provides the opportunity for networking in the beautiful city of Graz (Austria).

 

(C) RTDS – Portrait: Stephen Webb (RTDS), Martin Kavšček (ACIES BIO Ltd.), Nelo Emerencia (Director of Programming, Bio Based Industries Consortium BIC), Stefan Weiss (RTDS)

 

Presentations and Discussions on Trends in Science & Industry

Amongst 19 Sessions on “The learnings of H2020 and the Forecast to Horizon Europe”, “Novel Biorefinery and Biotechnology Concepts”, “Next Generation Biopolymers – Materials of the Future?” and 28 Science Flashes focusing on ideas with the potential to enter the market, SUSBIND was invited to the KETBIO Workshop: Boosting Biotech Research, were Stephen Webb (RTDS) presented the project and partake in the panel discussion with four other H2020 and innovative IB projects that encompass novel feedstocks and process technologies. Moreover, SUSBIND was presented over three days (November 18th-20th) in a twin booth with our Slovenian partners from ACIES BIO Ltd., who represented SUSFERT. Promoting SUSBIND and SUSFERT together helped disseminate both projects to a broader stakeholder group. Amongst the numerous visitors was Bio-based Industries Consortium (BIC) Programme Director Nelo Emerencia (Picture) and Ecoduna algae farm managing partner Dr. Silvia Fluch.

We are looking forward to the next esib in 2021, when Science meets Economy meets Politics again!

 

Author: Dr. Stefan Weiss

Dissemination & Communication / RTDS Group

SUSBIND Enables two PhD Dissertations with Wood K+

Within the scope of the EU-funded SUSBIND, in cooperation with different companies and university partners, our Austrian project partner Wood K plus provided two PhD student positions within the Area “Wood Materials Technologies” (WMT), where innovative adhesives are developed in the team “Advanced Bonding”.

The “Kompetenzzentrum Holz GmbH” (Wood K plus) provides research and development services for the industry in the field of wood composite materials and wood chemistry.

To read more news from Wood K plus go to: “News – Wood K plus”