HOTZYMES´s consortium has worked hard during this first year of the project to obtain promising results on the development of a Toolbox of Nanoparticles (NPs) with adequate and tunable magnetic heating efficiencies. This allows us to think that they could enable the development of the proposed concept to exert spatio-temporal remote control over enzymatic cascades. Besides, the partners had also been working on other aspects that are also key-enabling to achieve this aim. In this sense, they not only have been working on the development of the Toolbox of recombinant enzymes needed but also in the development of tailor-designed functionalization strategies for the obtained NPs and in the study of their safety.
During the second year of the project, HOTZMES´s consortium has shown the feasibility of triggering multiple local hotspots using a mixture of MNPs with different magnetic heating efficiencies. The results obtained are compatible with our aim of gaining spatio-temporal remote control over enzymatic cascades by the application of alternating magnetic fields (AMF). Besides, the partners have been working on the development of hybrids integrating MNPs and enzymes to avoid aggregation issues observed when using enzymes attached to MNPs directly on the biotransformations that were selected for exploring the feasibility of their AMF-tuning. In addition, the partners have been working on the development of different tools (recombinant protein toolkits, thermodynamic studies, nanothermometry measurements, etc) in order to help to understand the parameter governing and influencing the heat transferences processes triggered at the nanoscale by AMF.
During the final period of the project, partners have been working on obtaining a third generation of MNPs changing their anisotropy but maintaining their core size, also focusing on the development of more sustainable methods for their synthesis. Progress has also been made in modeling studies and the use of molecular dynamics to understand the heating at the nanoscale. A recombinant toolkit of enzymes has been developed that would allow studying the effect of enzyme orientation/distance to the MNP surface on heat transference processes triggered by AMF. The feasibility of using the selected enzymes to perform the selected bioprocesses was optimized. Integration of enzymes and MNPs within hybrid materials resulted in a suitable strategy to avoid MNP aggregation issues. Indeed, different AMF-tunable enzyme micro and nanohybrids were obtained. In addition, a pilot-scale AMF reactor was designed, prototyped, and manufactured. Lixiviation of metal ions upon AMF application and an original “omic” epigenetic study have been conducted to go beyond traditional nanotoxicity approaches. All this work has resulted in the identification of exploitable results including: i) self-standing scientific and technical knowledge providing results for further research activities, ii) development of products, and iii) development of results for their use in standardization activities. Within these exploitable results, a European patent has been granted (EP21382585.4). This patent could form part of the portfolio of an innovative SME company, for whose creation several actions have already been initiated such as: i) contacting investment funds, and ii) receiving coaching from Business Development Consultancies. Besides, another patent is being prepared. Although many results obtained could not be disseminated until this patent is filed, the consortium has produced so far 21 open-access publications that could also be found in HOTZYME´s Zenodo community.
