Construction of a FLEXIble and adaptable enZYMatic biotechnological platform for sustainablE industrial production of bio-based fatty amines from side stream materials

Project Information

FLEXIZYME

Grant agreement ID: 101157528

DOI

10.3030/101157528

EC signature date 17 May 2024

Start date 1 June 2024

End date 31 May 2028

Funded under

• Food, Bioeconomy Natural Resources, Agriculture and Environment

Total cost

€ 4 984 163,75

EU contribution

€ 4 984 163,75

Coordinated by

INSTITUTO TECNOLOGICO DEL EMBALAJE, TRANSPORTE Y LOGISTICA
Spain

Periodic Reporting for period 1 - FLEXIZYME (Construction of a FLEXIble and adaptable enZYMatic biotechnological platform for sustainablE industrial production of bio-based fatty amines from side stream materials)

Reporting period: 2024-06-01 to 2025-11-30

Summary of the context and overall objectives of the project

Fatty amines are key functional chemicals with cationic and surface‑active properties, broadly used in cosmetics, detergents, coatings, and agriculture. The global market is projected to reach USD 5.54 billion by 2027. Current industrial production relies almost exclusively on the fossil‑based “nitrile route,” which is energy‑intensive, generates hazardous by‑products, and requires harsh conditions and toxic catalysts. Selectivity limitations further reduce efficiency. Although enzymatic alternatives are emerging, their high cost, limited robustness, and fragmented development strategies hinder large‑scale adoption.
FLEXIZYME addresses these challenges by developing the first integrated biotechnological platform for producing bio‑based fatty amines (FA) from fat‑ and protein‑rich side streams.FLEXIZYME unites enzyme engineering, in‑silico design, data‑driven biocatalysis, advanced reactor concepts, and early downstream solutions into a systematic, cross‑disciplinary framework. Machine learning, structural modelling, and process control strategies support rapid development and optimisation.
The platform is guided by LCA and TEA to ensure environmental performance, cost‑competitiveness, and circularity. Industrial validation in the cosmetics, detergents, and agriculture sectors will confirm technology relevance and market readiness.
By valorising low‑value residues from the agro‑food and oleochemical industries, FLEXIZYME contributes directly to European Green Deal goals, the EU Bioeconomy Strategy, and the Circular Economy Action Plan. Target impacts include a 30% reduction in GHG emissions, FA production costs below €5/kg, and improved recovery efficiency compared to chemical benchmarks. Delivering a TRL5 platform, FLEXIZYME will support the emergence of new bio‑based value chains, influence consumer and market behaviour, and create new opportunities for the EU biotechnology and biochemical industry.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

During its first 18 months,FLEXIZYME established the complete scientific and technological foundation for a new enzymatic, circular and scalable route to FA. The project progressed from concept to a fully integrated, data‑driven platform covering feedstock valorisation, enzyme discovery, cascade biocatalysis, process modelling and early downstream development.
WP2 – Process design & control: Delivered the full conceptual process design, defined reactor configurations and mass/energy flows, and set up methodologies for dynamic modelling and early‑stage monitoring, including thermal imaging.
WP3 – Feedstock & building blocks: Characterised 28 lipid samples and three legumes, selecting coconut/palm distillates and lentils. Developed robust hydrolysis routes supplying consistent fatty acids and amino‑donor streams.
WP4 – In‑silico enzyme mining & engineering: Identified novel thermostable CAR families and 33 TA candidates using structural prediction and ML tools. Established engineering bases and improved expression of promising TAs.
WP5 – Enzyme screening & cascade set‑up: Screened CARs and TAs, selecting top catalysts. Built a functional ATP/NADPH recycling system (53% conversion) and demonstrated the first CAR–TA cascade for long‑chain substrates.
WP6 – Downstream & circularity: Developed initial purification strategies. Mapped valorisation routes for lentil‑derived side streams.
WP7 – Sustainability: Established LCA and TEA frameworks and completed first inventories, showing clear advantages over the fossil nitrile route.
Overall: A full feedstock‑to‑product architecture, validated enzymes, functional cascade system, and circularity and sustainability frameworks are now in place—positioning FLEXIZYME for optimisation, integration and scale‑up in the next phase.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

During the 1st RP, FLEXIZYME achieved several breakthroughs that clearly surpass current capabilities in FA production, enzyme engineering and circular bioprocessing. These advances establish the basis for a fully bio‑based, low‑energy and modular FA platform not yet available in industry.
A. A new biotechnological route for long‑chain fatty amines: FLEXIZYME demonstrated the first integrated CAR–TA enzymatic cascade capable of converting lipid and protein sidestreams into long‑chain FAs. Key achievements include validated CARs and TAs active on C12–C18 substrates, a fully functional ATP/NADPH recycling system, and the first proof‑of‑concept cascade with long‑chain fatty acids—marking a strategic departure from the fossil nitrile route.
B. Discovery of novel thermostable CAR families: Using in‑silico structural prediction, ML ranking and metagenomic mining, the project uncovered previously unknown CAR subclasses with shorter sequences, higher predicted thermostability (+15 °C) and unique domain architectures, significantly expanding the CAR enzyme landscape.
C. AI‑enabled enzyme engineering: Advanced AI workflows enabled catalytic mapping, thermostability scoring and PROSS‑based multi‑mutation design, producing the first soluble, high‑yield variants of challenging TAs such as FumITA. This represents major progress in engineering long‑chain TAs, a longstanding bottleneck.
D. Integrated process design and modelling: A full conceptual process architecture—unit map, early mass/energy balances, dynamic modelling tools and first downstream strategies—was completed far earlier than typical RIA timelines, positioning FLEXIZYME as a next‑generation biorefinery concept.
E. A fully circular value chain: The process valorises low‑value feedstocks and actively reuses side streams (glycerol, polysaccharides, xylan‑rich residues), demonstrating true systemic circularity across feedstock, reaction, purification and residual valorisation stages.