Today, the increase of the atmospheric CO2 originates from the utilisation of carbon fossil in our daily life. In addition to the development of alternative energy sources (solar, wind, nuclear), plant biomass is one of the main options to replace fuel for transportation sector. Thus, plant biomass is projected to play an important role in this European bioeconomy strategy. The second-generation biofuels relies on a cheap and abundant non-food material: the lignocellulose (LC). LC consists of a complex network of cellulose, hemicelluloses, lignin and proteins that cross-link with each other and is highly recalcitrant to chemical or biological degradation. However, this chemical complexity offers a vast potential in the development of biorefinery for renewable and sustainable molecules and materials for our daily lives. In Nature, lignocellulolytic microorganisms are able to metabolize and recycle plant-derived organic carbon. They achieve this by using complex arsenals of cell wall-degrading enzymes. Most of these enzymes display a modular architecture, composed of catalytic and non-catalytic modules. Some anaerobic bacterium produce a self-assembling multienzymatic complex anchored to the outer membrane that couple enzymes with complementary activities. Previous work has evidenced that spatial proximity is a key to the remarkable efficiency of the cellulosome but it is difficult to evaluate the effect of the distance and active site orientation on enzymatic synergism mainly because of the high flexibility of the cellulosome. Thus, controlling such spatial organization is of high importance to master enzymatic synergism and increase the efficiency yield of PCW deconstruction. To reach this goal, an original approach is required. CONCERTO propose to use the BioMolecular Welding tool, composed of two small proteins Jo and In that are able to spontaneously create an intramolecular isopeptidic bond. Once linked to each other, Jo-In is a rigid complex of around 6 nm wide, displaying available N- and C-terminus for fusion. Furthermore, the anti-parallel organization of Jo-In offers the ability to create chimeras and modulate the relative spatial organization of linked protein domains. However this technology is limited because there is only one pair of Jo-In, and no natural complementary pair exists. This limitation prevents from the development of more complex assemblies that are required to degrade LC. Therefore, in CONCERTO, we propose to tackle this limitation by developing new pair of Jo and In in order to create larger organization of multi-modular enzymes. Thanks to new pairs of Jo and In, original enzymatic complexes will be designed in CONCERTO and will be deeply investigating, on one hand by solving the structure of theses complexes in solution using SAXS analysis and on another hand by carefully characterising the product profile generated by such enzymatic complexes thanks to the development of an analytical strategy that does not exists today. We hypothesize that various spatial organization will alter the product profile of the hydrolysis of complex plant cell wall substrates, helping us to understand the catalytic activity/spatial organization/product profile relationship within nanomachine and paved the way toward the control of biomass deconstruction.

Replacement of fossils resources by renewable resources is essential to achieve a sustainable growth of human society. Ultimately, energy and carbon used to produce fuels and chemicals must be sourced directly from the Sun and CO2 and synthesized biologically. The CONCEPT project holds the promise by proposing a hybrid strategy, combining both chemical and biological routes, as a hybrid concept of converting CO2 into a value-added product using renewable energy to create an artificial carbon cycle. To achieve this challenge, a unique consortium is being assembled according to the key competences of its partners from different research disciplines needed to establish a novel production basis of complex molecules (more than C1) from CO2. As no single organization or country has the capacity to successfully bring about the innovations and demonstrations intended in CONCEPT on its own, a well-balanced consortium has been arranged, with partners being complementary to each other and willing to closely cooperate in creating a research environment as envisioned in CONCEPT’s ambitious aim. At the moment, 6 legal entities independent from each other (i.e. 4 academics, 2 SMEs) and from 4 different eligible countries (i.e. France, Germany, Spain, Poland) have accepted to be part of the consortium. In the future more legal entities (2 companies, 2 research organisation) from 2 more different countries may join the consortium. Considering that CONCEPT is a pioneer project given the high-risk research due to the novelty of the concept and the huge social, environmental and political implications. Considering the highly competitive funding program in the call H2020-FNR-13-2020: “Bio-based industries leading the way in turning carbon dioxide emissions into chemicals”. The core Consortium headed by Stéphanie Heux and Claire Dumon requests help from the French Research Agency (ANR) to financially support the assembly of this European Scientific Network. In particular, this help will allow: - to strengthen and expand the consortium - to reinforce the project position - to implement actions in order to suppress or reduce identified weaknesses - to organise at three consortium meetings: one for creating the bases for an interdisciplinary and intersectoral collaboration and prepare the first stage submission , the second in order to strengthen the European proposal in view of stage two submission and a third to finalize the second stage proposition. The ambition is to submit a pre-proposal in January 2020 and, if accepted, a full proposal in September 2020. We are strongly convinced that the chances to succeed will be significantly increased with the MRSEI financial support. Also, the CONCEPT‘s concretisation will reinforce the French position in the bio-based manufacturing and processing domain, using biological and renewable raw materials.

Methanol is an attractive low-cost substrate for biotechnology. Although methanol-based cell factories have been established, there are still major gaps in understanding methylotrophy itself. Thus, limiting their use at industrial scale. In methylotrophic yeasts like Komagataella phaffii, the entire methanol utilization (MUT) pathway is confined in peroxisomes, which is essential for its functioning. In FUNCEMM, we aim to elucidate the meaning and importance of such spatial organization to apply it in the establishment of synthetic methylotrophy. We will first re-target the entire MUT pathway to the cytoplasm of K. phaffii to understand which MUT pathway reactions depend on peroxisomal localization. Second, artificial organelles based on bacterial microcompartments will be built to investigate whether the type of compartment is necessary for methylotrophy. Finally, we will harness this knowledge to build an artificial organelle mimicking the natural methylotrophic peroxisome to design superior synthetic methylotrophic Escherichia coli, capable of efficient use of methanol. Functionality of the MUT pathway variants will be assessed in vitro using in cell NMR analysis together with kinetic models while the engineered strains will be studied at the system level by combining multi-omics approaches with stoichiometric models. In one word, FUNCEMM proposes to apply the System & Synthetic Biology approaches to methylotrophy for industrial biotechnology. FUNCEMM brings together leading European expertise in natural & synthetic methylotrophy and biotechnology and will generate fundamental knowledge for a methanol bioeconomy.

GLYCINE is one of the twenty-two genetically encoded amino acids required for protein synthesis in all living systems. Although not considered as an essential amino acid, it is of increasing interest in the nutraceutical, cosmetic and pharmaceutical industry with a production > 22000 tons in 2018 and a estimated compound annual growth rate (CGAR) of 6,8% between 2020 and 2030. Commercialized glycine is currently exclusively produced by chemical means from fossil precursors. The aim of the G-BIOFERM project is to establish the foreground of a glycine production by microbial fermentation from renewable carbon sources, as a green and economically viable alternative. While we recently established the proof of concept of glycine bioproduction routes, we identified three major bottlenecks that must be unlocked in order to make this biotechnological process of glycine industrially attractive and economically viable. These bottlenecks which will be addressed in this G-BIOFERM are (i) the low catalytic efficiency of a key enzyme whose is critically important to reach optimal yield from sugars, (II) the apparent lack of glycine exporter and (iii) the high toxicity of glycine to the microbial cells. These bottlenecks will be addressed by combining systems and synthetic approaches, involving expertise in enzyme modelling, metabolic and strains engineering and fermentation process, with the full commitment of young French SME whose business model is to sell exclusively bio-based food supplements to its customers. The G-BIOFERM is therefore strategic and in full adequacy with the axis H.7 of the AAPG22 call: “Bioéconomie, de la biomasse aux usages” and will also perfectly meet society's expectations in terms of carbon neutrality, with a clean green product in the spirit of the French industrial revival supporting its small and medium-sized enterprises..

ABCardionostics envisions a radical shift in atherosclerosis management by offering unprecedented fully human antibody (HuAb)-based personalized diagnosis and therapy modalities for at-risk patients. ABCardionostics strength lies in a unique panel of fully HuAbs, selected by cutting-edge in vivo phage display in an atheromatous model. Of particular relevance, will be HuAbs targeting culprit or protective phenotypes of the highly plastic and heterogenous monocyte/macrophage (Mo/MP) lineage. Described as the “oracle of health and disease” for many pathologies, from cancer to inflammatory and infectious diseases, the Mo/MPs are also key actors in the progression of atherosclerotic plaques. The ratio between pro- and anti-atherosclerotic Mo/MP subsets is decisive in plaque progression toward rupture. By using HuAbs identified in atherosclerotic plaques and selected on the basis of their specificity for human Mo/MP subsets and immunoregulatory action, the ABCardionostics project will serve the non-yet achieved aim of disease monitoring and reversing the unbalanced functional state toward a protective activity, revolutionizing the current atherosclerosis imaging and therapeutic approaches. ABCardionostics will ultimately provide an accurate snapshot of the arterial tree in at-risk patients by 3D multi-marker guided positron emission tomography (PET) imaging, using specific HuAbs labelled with different isotopes to profile Mo/MP phenotype diversity, combined with 3D multi-sequence magnetic resonance imaging (MRI) and time-resolved three-directional phase-contrast MRI (4D MR Flow) to extract anatomical and advanced hemodynamic parameters. This imaging-based phenotyping to optimally identify plaque subtypes is the foundation of ABCardionostics’ radically new immunotherapeutic paradigm to deliver the right drug at the right place in the right patient. Bispecific HuAb-based immunotherapeutics will be designed to combine one HuAb targeting extracellular or cellular biomarkers to precisely identify in situ the affected site and another HuAb that will trigger the shift in the balance between culprit and protective Mo/MP subsets (initially identified by 3D imaging). The diagnostic and sophisticated therapeutic arms synergize to address the unmet need of personalized atherosclerosis treatment. The imaging protocols will be useful for the diagnosis and identification of patients who will benefit of HuAb-based immunotherapies and to monitor such treatments in an inherently personalized manner. New valuable atherosclerosis progression markers will be identified concomitantly with HuAb development, allowing novel scoring systems for efficient staging of vulnerable patients. To translate this research into clinical practice, the ABCardionostics consortium will combine cutting-edge biotechnologies and Artificial Intelligence to close the knowledge gaps on plaque MoMP spatiotemporal dynamics and the lack of imaging data combined to therapy: pioneering in vivo and in vitro phage display approaches to identify relevant HuAbs (CNRSa), multispectral mapping and MacroScreen to unravel the functionomics of MoMP subsets in atherosclerosis (UM), unvisited in silico and biochip development to identify HuAb targets (MAbS, CNRSc), innovative HuAb bioengineering techniques (CNRSb,d, BTM), derivatization and labelling to develop PET tracers to explore a translational non-invasive multi-marker PET/ MRI system for diagnosis and monitoring (CNIC) and an adapted immunotherapy strategy targeted to the appropriate plaque by designing bispecific antibodies (CNRSb). Deep learning methods will integrate multimodal imaging data to obtain insight into plaque morphology and composition (PMI). ABCardionostics successful completion will bring HuAb leads for atherosclerosis theranosis, and novel tools for HuAb identification and design, easily transposable to other diseases, revolutionizing the current imaging and therapeutic approaches.