Workpackage 2: Enantiopure 1,3/1,4-amino acids and amino alcohols from prochiral ketoacids

Workpackage Leader: Jun.-Prof. Dr. Robert Kourist
Lead beneficiary: Ruhr University Bochum
Participating Partners: Entrechem, SARomics, DSM, Viazym
Start month: 7 End month: 42

Description of work and role of partners

WP2 focuses on the synthesis of enantiopure 1,3- and 1,4-aminoacids: ESR4 develops a two-enzyme cascade combining amino acid α- aminotransferase (AAT) and aminoacid hydroxylase (AAH) to convert readily available keto acid precursors into high-value products. The ‘smart cascade’ uses the side-product α-ketoglutarate of the first step as external cofactor for the second step. Further conversion to 1,3- and 1,4-aminoalcohols by aminoacid decarboxylases (AAD) is a novel route for the synthesis of these valuable building blocks. ESR5 focuses on expanding the substrate scope of amino acid hydroxylases in order to create a generally applicable three-step cascade. Focus lies on the comparison of different AADs. The ESR identifies potential key amino acids and uses the CASTing approach to vary these residues. Screening of mutant libraries is done by a novel high throughput assay. Establishment of the cascade takes place at the academic partners, application for the synthesis of industrially relevant building blocks and scale-up at DSM.

Objectives

    1.) Avoiding cross-reactivity by flow-chemistry.
    2.) Generation of a broad-spectrum amino acid decarboxylase by protein engineering.
    3.) Proof-of-principle of a new three-enzyme cascade with in situ cofactor generation.

The Biocascades Project is a joint collaboration among the following university and industrial partners