Periodic Reporting for period 1 - FuturEnzyme (Technologies of the Future for Low-Cost Enzymes for Environment-Friendly Products)
Berichtszeitraum: 2021-06-01 bis 2022-11-30
FuturEnzyme is a project funded by the European Union’s Horizon 2020 research and innovation programme (grant agreement No 101000327). The consortium gathers technical and technological capabilities of 8 universities/research institutions, 3 small and medium enterprises, 2 clusters, and 3 industries, from 7 countries. FuturEnzyme is a project that meets consumers', industries' and planets' demands. Consumers demands, because their concerns about the environment, carbon emissions, and everyday consumption choices. Industries demands because of their determination to keep pursuing a 100% sustainable model of production and consumption. Planet demands, because cheosphere & biosphere are changing fast (CO2 emissions: 36.2 Giga-tons (Gt) in 2022; +2.2˚C in 2100). Enzymes can contribute meeting these demands by decreasing annually CO2 emissions by 1-2.5 billion-tons. This is why the general objective of FuturEnzyme is to establish and combine a series of technologies to retrieve microbial enzymes with exquisite performance and stability and low cost. With them, we aim to approach the complex reality and challenges of real consumer products in the detergent, textile and cosmetic sectors, with the ambition of greening products already in the market, making them more environmentally friendly, valuable, functional and sustainable. Detergents, textiles and cosmetics are our target products because they are highly consumed (222 Mt/year) and contribute globally to carbon emissions (1400 Mt CO2/year). The FuturEnzyme concept to approach the bottlenecks for the target sectors and products is as follows. For detergent, developing new and improved enzymes, lipases/esterases, which help to remove fatty stains at low wash temperatures (20-30°C) give the consumers new opportunities to reduce the carbon footprint and the energy needed for heating the water during laundry. However, it is very challenging to a) identify new enzyme candidates, which bring new and promising properties, and b) to evaluate if these wild type enzymes are applicable for our target. For textile, the utilization of enzymes in the textile production unit can have an effective impact to save energy, time and water and reduce CO2 emissions in three steps, namely, the removal of the spinning oils during the solvent cleaning step (by lipases/esterases), the removal of the dyes after the dyeing/fixing of the textile materials, and the degradation of textile waste after the end-use (by esterases/lipases, oxidoreductases and poly-ester hydrolases). The textile’s complexity, in the base material used and different oils & dye recipes & variants in color, make it quite complex to find only one enzyme in their removal and discoloration and neutralization of the wastewater. For cosmetic, enzymes (hyaluronidases) can have an effective role in hydrolyzing the long hyaluronic acid in a short-defined molecule that is key to the production of anti-ageing cosmetics. However, it is not so easy to find an enzyme, that cleaves/hydrolyzes this complex molecule at the right place to have a repeatable defined size with highest anti-ageing properties. The complexity of the products to be implemented together with the bottlenecks in selecting the best enzymes from all the millions available in nature are the biggest challenges. Indeed, finding such new enzymes is costly in money (€30k/enzyme) and carbon footprint (0.008-0.38 kgCO2 eq/enzyme), time-consuming (15 months/enzyme), and their production is also contributing to carbon footprint (8.9 gCO2 eq/g enzyme).
FuturEnzyme proposes an integrated solution that, starting with a clear knowledge about the manufacturers' needs, specifications and priorities and the inventory of the detergent, textiles and cosmetic benchmark, allow a targeted search for the best enzymes using big biodata mining, disruptive machine learning, activity-based bio-prospecting, protein engineering, nano-biotechnology, upscale fermentation, and downstream processing systems. Then, analyzing technology, market, consumer, and socio-economic demands, and assessing safety, risk, and life-cycle analysis during the technology and product development in an iterative process, will ensure that the results will be effective and relevant. These activities constitute the FuturEnzyme WorkPlan, that also includes a strong dissemination, communication and exploitation strategy. The expected results of the project include 2 web-based resources to screen the target enzymes, 1,000 microbial enzymes with manufacturers’ requirements (180 validated, 18 to outlast the product conditions), 3 innovative consumer products at defined scales, and dissemination, communication and exploitation actions. Overall, main activities performed included an exhaustive compilation of the manufacturers' requirements, based on which a number of protocols were implemented to screen a large diversity of bio-resources from at least 293 different locations and more than 1 billion public sequences. As a result, we retrieved 1,612 new enzymes: 678 being successfully produced, 368 validated, and 27 prioritized. The inventory of the benchmarks, LCA data, preliminary analysis of consumers’ behavior and requirements for the pre-industrial fermentation and validation tests was completed, collected and shared. Communication and dissemination, data management and exploitation plans, and a plan to collaborate with other EU projects funded were implemented. Main future activities include strengthening our collaboration with the other funded projects, performing new rounds of refine enzyme bio-prospecting, engineering and production (already ongoing), producing appropriate amounts of priority enzymes to do the first pre-industrial validations, completing the impact assessment of the LCA methodology and continuing dissemination and communication actions in close connection to exploitation actions.
Results are progressing beyond the state of the art. It is worth mentioning: i) the establishment of the Cluster Enzymes for Greener Products; ii) technical requirements for three target consumer products in the market; iii) novel methods, algorithms and biocontainers for bio-prospecting, from one of the largest collections of bio-resources, the target enzymes; iv) multiple expression systems, including expression via in vitro, organo-catalysts and active site design; v) a heuristic algorithm, AsiteDesign, to design and redesign active sites, and a novel strategy for enzyme “fishing, immobilizing and shielding”; vi) an unique set of enzymes with high potential for product innovation; vii) real-life products to proceed with pre-industrial validations. To highlight also, the impact at the policy level (policy document in preparation), on dissemination, communication and exploitation of results (audience of +35,000 people), training (4 PhD/Master Thesis defended and 16 in progress), recruitment (15 persons), gender dimension (including evaluating consumers’ behavior by gender), internationalization activities through resource mobilization. Finally, the activities and objectives are still priority for the three industrial partners, which reinforce the positive economic and environmental impact of the project.