Development of a biotechnological enzyme cascade process for the production of optically pure beta amino acids

• Applicant: Dr.-Ing. Ulrike Engel
• Subject Area: Biological Process Engineering
• Term: from 2016 to 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 290767188

The objective of this project is the development of a process for the synthesis of chiral beta3-amino acids based on an enzymatic cascade reaction starting from (aryl-) substituted dihydropyrimidines. The enzyme cascade is to be consisting of a dihydropyrimidinase and an enzyme for the decarbamoylation of N-carbamoyl-beta3-amino acids. Thus, starting from educts which can be synthesized from simple precursors (urea and cinnamic acid derivatives), a direct access to chiral beta3-amino acids is provided.A variety of different dihydropyrimidinases and biocatalysts with dihydropyrimidinase activity is available for this project. Additionally a selectively screening for enzymes able to decarbamoylize beta3-amino acids is to be conducted for the first time. Until now no enzymes catalyzing this reaction are described. However, this is due to the fact the enzymatic decarbamoylation only recently shifted in the focus of biocatalytic research. It is expected that several enzymes with this ability are discovered within this project.In detail dihydropyrimidinases and decarbamoylating enzymes are to be identified with molecular biological techniques, extensively biochemically characterized, optimized and finally provided for the process as recombinant enzymes. These enzymes are to be equipped with affinity-tags suitable for the enzyme purification as well as for the immobilization in microfluidic systems.Furthermore the provided new (aryl-) substituted substrates are to be characterized in detail and appropriate analytical methods are to be established (HPLC, photometer, thin layer chromatography, polarimeter).With these results different new and innovative prozess types (the Teabag-system for the immobilization of whole cells and the immobilization of purified enzymes in a microfludic system) are to be tested and compared.At the end of the project a toolbox of different dihydropyrimidinases and decarbamoylating enzymes is available. Applying the optimal process type the desired chiral beta3-amino acids can be efficiently produced with the enzymatic cascade process.

• DFG Programme: Research Grants