Periodic Reporting for period 1 - EXPERIMENTAL (EXploring Photoinduced Enzyme pRomIscuity in the Glucose-Methanol-Choline oxidoreductase family to dEvelop New phoTocAtaLysts)
Periodo di rendicontazione: 2022-08-15 al 2024-08-14
The goal of this project is to develop new stereospecific chemical reactions using enzymes activated by light, known as photobiocatalysis. Photobiocatalysis combines the use of light with an enzyme to access new chemical reactions that are difficult or impossible to achieve with traditional methods. By using light to activate enzymes, this project aims to make chemical synthesis processes more efficient by reducing the number of steps needed and avoiding the use of harsh conditions. Enzymes are natural catalysts that operate under mild conditions and have a unique chiral environment, allowing them to selectively produce one of two mirror-image forms of a molecule (enantioselectivity). This makes them particularly useful for producing complex molecules, such as pharmaceuticals, which require high precision. The reactions developed in this project could replace traditional chemical processes, leading to more sustainable and environmentally friendly chemical production.
The project roadmap involves identifying enzymes that can catalyze reactions using light and optimizing these enzymes. By expanding the capabilities of enzymes like Fatty Acid Photodecarboxylase (FAP), GMC oxidoreductase and other enzymes such as lactate monooxygenase (LMO), the project aims to provide more efficient and sustainable synthetic pathways. The research contributes to the broader objective of enhancing the use of biocatalysis in chemical manufacturing, addressing the need for greener, more efficient production methods that align with EU sustainability goals.
One of the key aspects of this project is to explore enzyme promiscuity in the context of photochemical activation. Enzyme promiscuity refers to the ability of enzymes to catalyze reactions beyond their natural substrate range. By using light to activate these enzymes, we aim to unlock new reactivity that is not typically accessible under standard biological conditions. This approach not only expands the scope of enzyme-catalyzed transformations but also provides a means to discover new synthetic pathways that are more selective and environmentally benign.The project builds on the promising activity observed in enzymes such as FAP, which naturally catalyzes decarboxylation reactions under light activation. We have been able to develop radical addition reactions with FAP and LMO, which are valuable in creating carbon-carbon bonds, a critical step in many chemical syntheses.
The integration of photobiocatalysis into synthetic chemistry represents a significant advancement over conventional chemical methods. Traditional chemical synthesis often requires high temperatures, extreme pH, and toxic reagents, which contribute to environmental pollution and increased energy consumption. In contrast, light-activated enzymatic processes occur under mild conditions, reducing both the energy requirements and the environmental footprint of chemical production. This aligns well with the EU's commitment to sustainable industrial processes and the development of green technologies.
The impact of this project extends beyond the laboratory, as it aims to contribute to the development of sustainable manufacturing processes. By demonstrating the feasibility of using light-activated enzymes for complex chemical syntheses, we hope to encourage the adoption of biocatalysis in the pharmaceutical and chemical industries. This would not only reduce the reliance on traditional, energy-intensive chemical processes but also provide a pathway for the production of high-value compounds in a more sustainable and cost-effective manner.
The project roadmap involves identifying enzymes that can catalyze reactions using light and optimizing these enzymes. By expanding the capabilities of enzymes like Fatty Acid Photodecarboxylase (FAP), GMC oxidoreductase and other enzymes such as lactate monooxygenase (LMO), the project aims to provide more efficient and sustainable synthetic pathways. The research contributes to the broader objective of enhancing the use of biocatalysis in chemical manufacturing, addressing the need for greener, more efficient production methods that align with EU sustainability goals.
One of the key aspects of this project is to explore enzyme promiscuity in the context of photochemical activation. Enzyme promiscuity refers to the ability of enzymes to catalyze reactions beyond their natural substrate range. By using light to activate these enzymes, we aim to unlock new reactivity that is not typically accessible under standard biological conditions. This approach not only expands the scope of enzyme-catalyzed transformations but also provides a means to discover new synthetic pathways that are more selective and environmentally benign.The project builds on the promising activity observed in enzymes such as FAP, which naturally catalyzes decarboxylation reactions under light activation. We have been able to develop radical addition reactions with FAP and LMO, which are valuable in creating carbon-carbon bonds, a critical step in many chemical syntheses.
The integration of photobiocatalysis into synthetic chemistry represents a significant advancement over conventional chemical methods. Traditional chemical synthesis often requires high temperatures, extreme pH, and toxic reagents, which contribute to environmental pollution and increased energy consumption. In contrast, light-activated enzymatic processes occur under mild conditions, reducing both the energy requirements and the environmental footprint of chemical production. This aligns well with the EU's commitment to sustainable industrial processes and the development of green technologies.
The impact of this project extends beyond the laboratory, as it aims to contribute to the development of sustainable manufacturing processes. By demonstrating the feasibility of using light-activated enzymes for complex chemical syntheses, we hope to encourage the adoption of biocatalysis in the pharmaceutical and chemical industries. This would not only reduce the reliance on traditional, energy-intensive chemical processes but also provide a pathway for the production of high-value compounds in a more sustainable and cost-effective manner.
The project initialy focused on screening enzymes within the Glucose-Methanol-Choline (GMC) oxidoreductase family for new photochemical reactions. Initial challenges in establishing a screening process were overcome, and significant progress was made with the identification of FAP as a promising enzyme for light-driven reactions.
The identified reaction involved radical addition after decarboxylation of phenylacetic acid, reacting with methyl vinyl pyridine. Similar reactivity was also observed with LMO, which showed better efficiency and fewer by-products. This led to a focus on evolving LMO for industrial applications, successfully increasing reaction yields from 10% to 30%.
The relocation of laboratory facilities led to some delays. However, this time was effectively used to draft articles, supervise students, and submit research proposals. Notably, a PhD student was able to complete a publication on the microbial photoproduction of hydrocarbons using FAP, currently available on bioRxiv.
Key scientific achievements include publications detailing the catalytic mechanism of FAP, the asymmetric synthesis of α-chloroamides using Ene reductase from the Old Yellow Enzymes familly, and a review on photobiocatalytic strategies, contributing significantly to the understanding of enzyme-driven photochemical processes.
The identified reaction involved radical addition after decarboxylation of phenylacetic acid, reacting with methyl vinyl pyridine. Similar reactivity was also observed with LMO, which showed better efficiency and fewer by-products. This led to a focus on evolving LMO for industrial applications, successfully increasing reaction yields from 10% to 30%.
The relocation of laboratory facilities led to some delays. However, this time was effectively used to draft articles, supervise students, and submit research proposals. Notably, a PhD student was able to complete a publication on the microbial photoproduction of hydrocarbons using FAP, currently available on bioRxiv.
Key scientific achievements include publications detailing the catalytic mechanism of FAP, the asymmetric synthesis of α-chloroamides using Ene reductase from the Old Yellow Enzymes familly, and a review on photobiocatalytic strategies, contributing significantly to the understanding of enzyme-driven photochemical processes.
The project expanded the scope of enzyme promiscuity by developing novel photoenzymatic pathways. The improvements made to LMO mutants resulted in increased catalytic efficiency, which is crucial for making these enzymes more useful in industrial settings.
The work on FAP and LMO demonstrated that light can be used to activate complex reactions involving radical intermediates, providing an innovative way to create complex molecules under mild conditions. This offers a sustainable alternative to traditional chemical synthesis, with fewer steps and reduced energy consumption.
Future work will aim to broaden the substrate scope of these enzymes and further optimize their catalytic properties for industrial applications. Funding is being sought to acquire dedicated robotic screening platforms to enhance the throughput of enzyme optimization and reaction discovery. The goal is to make photobiocatalysis a practical and scalable solution for industrial synthesis, particularly for producing enantiomerically pure compounds.
The work on FAP and LMO demonstrated that light can be used to activate complex reactions involving radical intermediates, providing an innovative way to create complex molecules under mild conditions. This offers a sustainable alternative to traditional chemical synthesis, with fewer steps and reduced energy consumption.
Future work will aim to broaden the substrate scope of these enzymes and further optimize their catalytic properties for industrial applications. Funding is being sought to acquire dedicated robotic screening platforms to enhance the throughput of enzyme optimization and reaction discovery. The goal is to make photobiocatalysis a practical and scalable solution for industrial synthesis, particularly for producing enantiomerically pure compounds.