Periodic Reporting for period 2 - RADICALZ (RAPID DISCOVERY AND DEVELOPMENT OF ENZYMES FOR NOVEL AND GREENER CONSUMER PRODUCTS)
Reporting period: 2022-12-01 to 2024-05-31
The current climate and sustainability crises emphasize the need for an accelerated transition towards a circular bioeconomy as part of a greener and more competitive European industry. The application of enzymes in industrial processes is increasingly important to achieve the EU’s sustainability goals and strengthen the circular bioeconomy, because of their capacity to transform (bio)molecules selectively and efficiently. Thus, enzymes can replace oil-based chemistry for the use bio-based feedstocks, preferably from secondary waste streams, transitioning from linear to circular consumption models.
However, enzymes still find hurdles for their industrial application: low success rates of discovery and engineering; tedious and expensive methods to explore diversity; and limited activity/stability in the final application. RADICALZ assembles an interdisciplinary and intersectoral consortium to deliver faster, more versatile and more affordable tools for enzyme discovery and engineering, enabling the development of novel enzymes, new formulations and ingredients for more environment-friendly and healthier consumer products in a timeframe compatible with industrial development. To this end, RADICALZ is: i) developing new droplet microfluidic tools to find suitable enzymes for consumer products; ii) developing user-friendly software solutions based on machine learning (ML) for faster and more accurate enzyme engineering; iii) developing novel enzymes and bio-based, bio-catalytically synthesized ingredients for consumer products (glycosides, wash-enhancing enzymes, bio-based thickeners, natural antioxidants and fragrances); iv) developing bio-based, condition-responsive capsules for the protection and triggered release of enzymes and ingredients in the formulation of consumer products.
However, enzymes still find hurdles for their industrial application: low success rates of discovery and engineering; tedious and expensive methods to explore diversity; and limited activity/stability in the final application. RADICALZ assembles an interdisciplinary and intersectoral consortium to deliver faster, more versatile and more affordable tools for enzyme discovery and engineering, enabling the development of novel enzymes, new formulations and ingredients for more environment-friendly and healthier consumer products in a timeframe compatible with industrial development. To this end, RADICALZ is: i) developing new droplet microfluidic tools to find suitable enzymes for consumer products; ii) developing user-friendly software solutions based on machine learning (ML) for faster and more accurate enzyme engineering; iii) developing novel enzymes and bio-based, bio-catalytically synthesized ingredients for consumer products (glycosides, wash-enhancing enzymes, bio-based thickeners, natural antioxidants and fragrances); iv) developing bio-based, condition-responsive capsules for the protection and triggered release of enzymes and ingredients in the formulation of consumer products.
In RP1 and RP2, RADICALZ partners have selected candidate enzymes and evolved them for their use as ingredients in “smart” consumer products as well as for syntheses of bio-based ingredients for laundry detergents, nutraceuticals and cosmetics. The Consortium’s vast resources, including large collections, metagenomes acquired in past EU-funded actions, private and public databases, continue to constitute a priceless asset for novelty, esp. when combined with in silico or in vitro mining of sequence space, such as the proprietary 3DM systems of BIOP, sequence similarity network (SSN)-based mining at BRAIN and INSAT or the microfluidic workflows developed at UNEXE, UAM and INSAT. In figures, RADICALZ set out to discover and engineer enzymes for 10 reactions or applications, fulfilling this objective at 100%. Moreover, the biocatalytic production of 4 additional natural ingredients for consumer products and 2 additional applications of microfluidics-based enzyme formulation in consumer products are being undertaken.
Technology is one of the trademarks of RADICALZ, putting forth ML and microfluidics to speed up enzyme discovery and engineering. In this respect, RADICALZ has developed 4 new droplet assays, including 2 in the last reporting period. The ML tools are being developed on schedule and their most recent iterations keep outperforming the state of the art. Moreover, they are beginning to profit from experimental training data obtained in the enzyme engineering campaigns and vice versa in a virtuous cycle.
Promoting a “culture of metrics” in biocatalysis is another trademark of RADICALZ. Process metrics and modelling are being used project-wide to pinpoint potential pitfalls of the envisioned enzymatic processes. The former has outlined the critical relevance of process intensification and the process of raw stuff extraction (upstream) as well as wastewater treatment after the reaction (downstream). For one of the processes, a complete LCA has already been delivered. Moreover, this culture of metrics is being promoted beyond the life of the project through a series of publications by partner SUSMOM (see Cordis). Regarding the latter, three different processes have been modelled in detail, uncovering substrate and product inhibitions that will result in the substitution of enzymes or changes in the substrate feeding regimes. Therefore, we have illustrated the general validity of the diagnostic tools of RADICALZ, similarly to the enzyme discovery tools. Finally, targeted and timely innovation and exploitation analyses in tandem with awareness raising via advanced communication tools maximize the impact of the project, which at this point includes several publications, 1 patent, numerous outreach and dissemination activities a newsletter and a policy brief.
Public deliverables can be accessed in Cordis. Public summaries of confidential deliverables and publications can be found at https://zenodo.org/communities/radicalz(opens in new window).
Technology is one of the trademarks of RADICALZ, putting forth ML and microfluidics to speed up enzyme discovery and engineering. In this respect, RADICALZ has developed 4 new droplet assays, including 2 in the last reporting period. The ML tools are being developed on schedule and their most recent iterations keep outperforming the state of the art. Moreover, they are beginning to profit from experimental training data obtained in the enzyme engineering campaigns and vice versa in a virtuous cycle.
Promoting a “culture of metrics” in biocatalysis is another trademark of RADICALZ. Process metrics and modelling are being used project-wide to pinpoint potential pitfalls of the envisioned enzymatic processes. The former has outlined the critical relevance of process intensification and the process of raw stuff extraction (upstream) as well as wastewater treatment after the reaction (downstream). For one of the processes, a complete LCA has already been delivered. Moreover, this culture of metrics is being promoted beyond the life of the project through a series of publications by partner SUSMOM (see Cordis). Regarding the latter, three different processes have been modelled in detail, uncovering substrate and product inhibitions that will result in the substitution of enzymes or changes in the substrate feeding regimes. Therefore, we have illustrated the general validity of the diagnostic tools of RADICALZ, similarly to the enzyme discovery tools. Finally, targeted and timely innovation and exploitation analyses in tandem with awareness raising via advanced communication tools maximize the impact of the project, which at this point includes several publications, 1 patent, numerous outreach and dissemination activities a newsletter and a policy brief.
Public deliverables can be accessed in Cordis. Public summaries of confidential deliverables and publications can be found at https://zenodo.org/communities/radicalz(opens in new window).
The tools, enzymes, processes and molecules developed in the first and second reporting periods and those under ongoing development build on previous developments of RADICALZ partners, who represent the state-of-the art in their respective disciplines. As a non-comprehensive summary, RADICALZ has gone beyond the state of the art and created impact by:
• Developing 4 novel enzyme assays and 2 workflows in droplet format to speed up enzyme discovery and reduce assay costs. In RP3, these will be used to engineer enzymes and generate training data for ML. These will result in the reduction of costs to develop/discover novel enzymes, in turn impacting on the reduction of costs of the biocatalytically synthesized molecules and ultimately of consumer products.
• Developing innovative ML tools, which build on previous know-how of partner BIOP and which keep outperforming all benchmarked tools. In RP3, further evolution of these tools is planned by incorporating experimental training data. These tools are the seed of AI-guided protein engineering, to increase the efficiency, reduce the time needed to improve proteins for industrial application, and ultimately democratize protein engineering.
• demonstrating the feasibility of previously undescribed enzymatic reactions, e.g.: several glycosylations and aqueous acylation reactions, including the modification of bulky molecules and polyphenols. In RP3, effort will concentrate towards reaction optimization. This will result in greener, more sustainable consumer products, whose ingredients are biosourced, thus contributing to sustainability and a circular bioeconomy
• analysing the economics but also the sustainability of reactions, considering upstream, reaction itself and the downstream of case study reactions. Although some of these recommendations are reaction specific, others are generalistic (extraction of substrates, wastewater treatment, intensification and therefore constitute a roadmap for process development that may feed policy recommendations in the second policy brief with the sibling projects.
• optimizing the communication and dissemination of the project over social media, including expanding the range of stakeholders that the project can reach. In RP3, we expect to keep the continued growth shown in RP2 with a focus towards dissemination of results and engagement of industrial stakeholders. Ultimately, the use of targeted dissemination results in enhanced impact of the project, maximising the return on investment in research and innovation, and promoting the effort of the European Commission towards a resilient, competitive and sustainable European economy.
• Developing 4 novel enzyme assays and 2 workflows in droplet format to speed up enzyme discovery and reduce assay costs. In RP3, these will be used to engineer enzymes and generate training data for ML. These will result in the reduction of costs to develop/discover novel enzymes, in turn impacting on the reduction of costs of the biocatalytically synthesized molecules and ultimately of consumer products.
• Developing innovative ML tools, which build on previous know-how of partner BIOP and which keep outperforming all benchmarked tools. In RP3, further evolution of these tools is planned by incorporating experimental training data. These tools are the seed of AI-guided protein engineering, to increase the efficiency, reduce the time needed to improve proteins for industrial application, and ultimately democratize protein engineering.
• demonstrating the feasibility of previously undescribed enzymatic reactions, e.g.: several glycosylations and aqueous acylation reactions, including the modification of bulky molecules and polyphenols. In RP3, effort will concentrate towards reaction optimization. This will result in greener, more sustainable consumer products, whose ingredients are biosourced, thus contributing to sustainability and a circular bioeconomy
• analysing the economics but also the sustainability of reactions, considering upstream, reaction itself and the downstream of case study reactions. Although some of these recommendations are reaction specific, others are generalistic (extraction of substrates, wastewater treatment, intensification and therefore constitute a roadmap for process development that may feed policy recommendations in the second policy brief with the sibling projects.
• optimizing the communication and dissemination of the project over social media, including expanding the range of stakeholders that the project can reach. In RP3, we expect to keep the continued growth shown in RP2 with a focus towards dissemination of results and engagement of industrial stakeholders. Ultimately, the use of targeted dissemination results in enhanced impact of the project, maximising the return on investment in research and innovation, and promoting the effort of the European Commission towards a resilient, competitive and sustainable European economy.