TOPCAPI will exploit the natural fabrication power of actinomycetes as microbial cell factories to produce three high value compounds: GE2270, a starter compound for the semi-synthesis of NAI-Acne, a new topical anti-acne drug in Phase II clinical trials; 6-desmethyl-tetracycline (6DM-TC) and 6-desmethyl 6-deshydro tetracycline (6DM6DH-TC), intermediates for semi-synthetic conversion to medically important type II polyketide tetracyclines (TC), e.g. minocycline, tigecycline, and the novel omadacycline, which is in Phase III clinical trials, to be used against Methicillin-resistant Staphylococcus aureus infections. Our work will focus on two bacterial host species: Streptomyces coelicolor and Streptomyces rimosus. These host species will be characterised using systems biology approaches, applying integrated data analysis to transcriptomics and metabolomics experiments, combined with predictive mathematical modelling to drive the rapid improvement of these microbial cell factories for industrial drug production using advanced metabolic and biosynthetic engineering approaches. At the same time, we will establish an expanded toolbox for the engineering of actinomycete bacteria as cell factories for other high added-value compounds. In the proposed 4-year project, we will: 1. Host engineer two new actinomycete strains for industry-level improved heterologous compound production through integrating systems biology-driven strain design and state-of-the-art genome editing. 2. Engineer the biosynthesis pathways to obtain high-efficiency synthesis of GE2270 and new pathway variants for 6DM-TC and 6DM6DH-TC as well as improve its production purity. 3. Optimise the expression of the engineered target pathways in pre-engineered strains to achieve industrially viable production levels of ≈1 g/L for GE2270 and ≈24 g/L for 6DM-TC, while creating a complete novel production strain for 6DM6DH-TC.

ACIES BIO has developed an innovative and disruptive high-value technology to address a major economical and environmental challenge of the world’s dairy industry: waste whey. Over 200 million tons of whey is generated annually, and only limited economical solutions exist to process it. The innovative patent pending technology Whey2Value uses a unique bioprocess to utilize whey as a primary ingredient for microbial fermentation to produce sustainable high-value products, such as vitamin B12. The technology greatly reduces the negative impact on the environment by almost eliminating the organic content of the wastewater, allowing for its recycling, while the product of the technology is a protein-rich biomass with high content of vitamin B12 to be used as a very high quality animal feed to complete the dairy industry’s circle. The innovation W2V is perfect example of how circular economy should work. The Whey2Value technology requires a very low-cost processing facility, which can be installed on site, and minimal maintenance costs. It represents a truly unique opportunity to create a huge and disruptive impact on dairy industry, particularly for the competitiveness of small and medium sized European dairy companies, generating high-value products from waste material, and at the same time creating a sustainable solution with a greatly reduced burden to the environment. The objective of this project proposal is to prepare a thorough business plan and feasibility study in the scope of Phase 1, followed by scaling-up and demonstration of operational technology in industrial setting with a local dairy company in the scope of Phase 2. The technology is ready for industrial demonstration, which will be followed by EU and global commercialization of Whey2Value. We expect a rapid worldwide market adoption of this disruptive eco-biotechnology.

Post-translational chemical modifications to histone proteins – the storage units of DNA – influence gene expression electrostatically and through specific protein-protein interactions, the study of which is known as epigenetics. Although many enzymes have been identified which add, remove or recognise these modifications, the implication of metabolism in the induction of epigenetic states is a recent development – particularly highlighting acetyl coenzyme A (CoA) as the sole donor for acetylation. Due to the lack of appropriate biochemical tools, this emerging field has not yet been exploited, however Acies Bio’s leading work on the efficient synthesis of 4’ phosphopantetheine (4-PPT), a natural precursor of the prolific substrate CoA, places us in a unique position to realise the novel approach of metabolically inducing epigenetic modifications. In progress towards therapeutic use bypassing faulty metabolism in a rare genetic disorder, we have shown that 4-PPT is cell permeable, serum-stable, tolerated at high doses without side-effects, and can also upregulate histone acetylation. This presents a previously unexplored route to develop 4-PPT as a novel vehicle for the delivery of diverse intracellular acyl-CoA species, and subsequent histone modification. Taking advantage of the interplay between metabolic and epigenetic states, and the complementarity of backgrounds that Acies Bio and Dr Sekirnik can provide through an international partnership combining expertise in both fields, our proposed work would illuminate recently characterised histone acylations. By synthesising novel 4-PPT derivatives, and demonstrating their activity on a cellular and model organism level, we will develop new tools to enable rapid enrichment of rare endogenous modifications to probe their function. This would form the first demonstration of use of native enzyme substrates to affect epigenetic states, affording new European research opportunities in both metabolomics and epigenetics.

SUSFERT addresses the massive usage of mineral fertilisers in EU agriculture, which are largely based on non-renewable resources, but are required in intensive crop production for meeting demands for food and feed. SUSFERT will develop multifunctional fertilisers for phosphorus (P) and iron (Fe) supply, which will fit into existing production processes and common EU agricultural practice. The SUSFERT innovation combines bio-based and biodegradable coatings, probiotics, the renewable P-source struvite, and nutrigels in at least four novel sustainable P and Fe fertiliser products, which may partly or fully replace unsustainable and resource-intense conventional ones. SUSFERT will test novel organic formulations, microgranules, granules, soil improvers, nutrigels and liquid fertilizer products in field trials, evaluate them regarding economic potential and environmental sustainability, ensure regulatory compliance and establish industry-scale production processes (TRL 7-8). Technological concepts include 1) probiotics based on P and Fe solubilizing Bacilli and Actinobacteria; 2) microbial siderophore (Fe chelator) produced in a demonstration plant; 3) enzymatic modification of the by-product lignin for cost effective, biodegradable controlled release coatings and product stabilization and 4) demonstration of struvite, a renewable P-source from wastewater, as a partial substitute of mineral P. Five industrial, four SME and three academic partners will contribute expertise along the whole value chain in biotechnology, microbiology, large-scale fermentation, fertiliser production and sales. We will fit the SUSFERT products into the regulatory and policy context for conventional and organic agriculture in various pedo-climatic conditions in Europe and prepare rapid market entry post-project.