Periodic Reporting for period 1 - PIEZOZYMES (Enzymatic Piezoelectric Composites To Regenerate Redox-Cofactors Driven By Mechanical Sources.)
Periodo di rendicontazione: 2022-06-01 al 2024-11-30
One of the current challenges of modern biotechnology includes the cost-effective production of sustainably sourced biochemicals and biofuels. Redox-biosynthetic pathways are key catalytic tools in chemical manufacturing for creating a bio-based economy. A significant hurdle is the sustainable regeneration and recycling of redox cofactors, necessary for effective industrial use of many redox-biosynthetic pathways. This project seeks to overcome these challenges by developing new biotechnological tools for efficient redox cofactor regeneration using mechanical sources. This innovative approach offers significant advantages over current systems that rely on biochemical, electrochemical, and photonic methods. Utilizing mechanical energy for cofactor regeneration is anticipated to yield more robust, scalable, and cost-effective solutions for industrial applications. The progress from this project will revolutionize the field, enabling the use of cofactor-dependent enzymes as economically viable biocatalysts. This technology can transform various industrial biotransformations, making them more sustainable and economically feasible. Beyond technical advancements, the project will have a substantial social impact, opening new pathways for sustainable industrial practices, reducing reliance on non-renewable resources, and supporting a greener, bio-based economy.
During the first 24 months of the project, we have successfully completed the initial two work packages outlined for this period. Our efforts have led to the heterologous expression and purification of the necessary enzymes, laying a strong foundation for the project's subsequent phases. In addition, we have engineered a diverse array of active enzyme mutants by strategically introducing specific residues into the protein structure to facilitate their oriented immobilization. Our steady-state kinetic characterization of both the native enzymes and their variants revealed significant insights into their activity profiles. We complemented this with comprehensive electrochemical characterization on gold electrodes, which provided critical data on their electroactive properties. By meticulously controlling the orientation of the enzymes on the electroactive surface, we achieved a substantial enhancement in the output catalytic current, demonstrating the efficacy of our approach. These achievements not only validate our methodological strategies but also pave the way for future innovations in enzyme-based applications. The improved output catalytic current underscores the potential for these immobilized enzymes to be used in high-efficiency biocatalytic systems. We have also been synthesizing a diverse array of piezoelectric platforms to immobilize enzymes while controlling their morphology and functionality. Additionally, we are developing various activation chemistries to anchor enzymes onto these platforms effectively. Moving forward, these findings will be instrumental in guiding the next phases of our project, ultimately driving advancements in sustainable and efficient industrial processes.
Our novel approaches to redox cofactor regeneration are poised to make substantial contributions to the scientific community, particularly in the fields of biocatalysis and synthetic chemistry. By enhancing the efficiency and sustainability of the redox processes, these cutting-edge catalytic tools will revolutionize the use of oxidoreductases in large-scale applications. This advancement will not only improve the cost-effectiveness and environmental footprint of industrial processes but also enable new pathways for complex chemical synthesis and biotransformation. As a result, our work promises to open up new avenues for research and industrial application, driving innovation and fostering sustainable development in chemical manufacturing.