Project Details
Projekt
Establishing a new platform for H2-driven asymmetric catalysis by design of a fusion-enzyme
Subject Area
Biological Process Engineering
Term
since 2025
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 564665679
The aim of this research project is to establish a new platform for H2-driven asymmetric catalysis, applicable in the synthesis of fine chemicals. To realize this, an unprecedented fusion protein between the soluble [NiFe] hydrogenase (SH) from Cupriavidus necator and the Old Yellow Enzyme (OYE) ene-reductase from Thermus scotoductus will be designed. In previous studies, the OYE enzyme showed an enantiomeric excess ee > 99% for the asymmetric reduction of 2-methylmaleimide to (R)-2-methylsuccinimide, which could serve as precursor for antimicrobial substances. [1] The engineered fusion enzyme will allow biocatalysis with a high atom efficiency using H2 to deliver both electrons and protons for the asymmetric reduction. The investigation of the electron transfer in the fusion protein is one major goal in this research project. Therefore, different protein linkers with variable properties are applied to test the capability of a direct electron transfer. The latter would circumvent the need for expensive cofactor regeneration in biotransformations. In comparison to direct electron transfer, natural and artificial electron carriers are investigated. Characterization of kinetic and thermodynamic parameters as well as optimization of operational conditions will be conducted in part by using an automated reaction platform. Inline Fourier-transform infrared (FTIR) spectroscopy will enable quantitative substrate-product analysis without sampling, and chiral gas chromatography (GC) will validate the stereochemistry. Based on the inline analytics the development of an automated reaction platform integrated with a comprehensive research data management (RDM) system, incorporating data exchange standards like EnzymeML and contribution to databases such as BRENDA to ensure adherence to FAIR principles
DFG Programme
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