12-month Periodic Report Summary

Project context and objectives

Large-scale industrial production of chemicals often also produces large quantities of toxic waste. Improved biocatalysts for carbon-carbon bond construction will lead to novel safe industrial processes, for the benefit of industry, consumers and the environment.

Products of industrial organic synthesissuch as pharmaceuticals and bulk chemicals contribute significantly to the quality of life and the economic development of modern societies. However, industrial chemical processes carried out through “classic” chemistry and catalysis often involve the use of hazardous substances and considerable amounts of energy. In contrast, enzymes are natural catalysts that speed up the manufacture and modification of molecules under mild reaction conditions and with exquisite selectivity, leading to increased product yields. Thus, the application of enzymes (“biocatalysis”) to catalyze industrial reactions offers an important potential to improve chemical manufacturing processes by reducing the use of corrosive chemicals, organic solvents, toxic metals and energy, in compliance with the concept and principles of Green Chemistry.

Carbon-carbon bond forming reactions are key processes in industrial organic synthesis to construct advanced molecules of high value, but remain insufficiently explored due to the lack of robust enzymes that cover a sufficiently wide range of reactions. To bridge the gap between laboratory experiments and industrial large-scale production,the scientists behind CarbaZymes address these challenges by developing a broad toolbox of carbon-carbon bond forming enzymes and by improving their properties so that these enzymes can be applied in new sustainableindustrial processes with reduced environmental footprints.The CarbaZymes project will pursue the biocatalytic synthesis of several valuable compounds, spanning a range of four chiral fine chemicals and three bulk chemicals, corresponding to market needs detected by the industrial partners in the consortium.

Work performed and main results

Within the first year, the consortium has successfully established advanced tools to identify and develop a broad panel of improved enzymes for carbon-carbon bond formation in a supra-disciplinary manner. The toolbox of methodologies includes the construction of bioinformatics tools, tests for high-throughput reaction analysis, protein structure modeling, as well as fast protocols for mutagenesis and stabilization of enzymes. Microreactor technology has been set up for bioprocess characterization and advanced reaction engineering.

A bioinformatics information system was set up to facilitate an in-depth data mining for the several target enzyme superfamilies to aid in the discovery and development of novel biocatalysts. Structural models were built to support a deeper understanding of structure-function relationships within the known natural variety. Tools could be advanced that allow increasing the stability of a given enzyme at higher temperatures.

A first superfamily library of novel enzymes together with a collection of known enzymes were produced and subsequently tested for their performance in various reactions within the scope of the project. Several key enzymes could be identified that show promising properties for further development and for assembly into efficient catalytic reaction cascades. The first libraries of mutant enzymes were constructed in a search for catalysts having improved properties for the target reactions by in-vitro-evolution and high-throughput screening. A novel total synthesis protocol was achieved for the integrated synthesis of an amino acid precursor with high volumetric productivity.

A broad program was devised and started for public dissemination of results, including original scientific publications, workshops and practical training courses, as well as promotion of white biotechnology via contacts to the general public through internet presence, newsletter and social media channels.

Potential impact

The CarbaZymes outcomes will have both social and economic impact, by responding timely and efficiently to pressurizing market needs from an increasing and aging population, by addressing markets worth billions and creating new intellectual property. CarbaZymes is expected to promote innovation in the field of carbon-carbon bond formation at large scale, by developing natural enzyme-catalyzed routes to important specialty and bulk chemical precursors. The range of novel and improved biocatalysts will also provide opportunities for the synthesis of many further valuable products, thus having a profound impact on reducing emissions, energy consumption and toxic waste. By developing new intellectual property as well as advancing biotechnology know-how — a Key Enabling Technology for Europe — this project aims to strengthen the global competitiveness of the European chemical and pharmaceutical industry.

 

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