Producing chiral amines

a bi-enzymatic cascade involving Old Yellow Enzymes and amine dehydrogenases

Master thesis (2020)

Authors

E.P.J. Jongkind BT/Biocatalysis - AS

Contributors

C.E. Paul BT/Biocatalysis - AS (mentor)
U. Hanefeld BT/Biocatalysis - AS (mentor)
Adrie J.J. Straathof BT/Bioprocess Engineering - AS (mentor)
Research Group
BT/Biocatalysis (AS) (TU Delft), BT/Biocatalysis (AS) (TU Delft)
Copyright: © 2020 Ewald Jongkind
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Published Date

24-04-2020

Language

English

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Faculty

Applied Sciences

Department

BT/Biotechnologie

Research Group

BT/Biocatalysis

Abstract

Chiral amines are valuable compounds for their use as building blocks for pharmaceutical and fine chemical industries. Previously, chiral amines were made with metals as catalysts, but these are unsustainable and difficult to remove from the product. Organocatalysis is a sustainable alternative, but requires chiral ligands, which are costly. Over the last decade, biocatalytic methods have been developed as well. Our goal was to produce chiral amines with our designed bi-enzymatic cascades, containing an ene reductase and amine dehydrogenase. We produced and purified ene reductases and assayed the asymmetric reduction of our proposed substrate scope, consisting of unsaturated ketones and aldehydes. We also investigated the impact of multiple reaction conditions on these performances that are optimal for amine dehydrogenases. Starting with 10 mM of substrate, we obtained concentrations up to 9.7 mM amine. Also, we obtained 3-methylcyclohexylamine with an enantiomeric and diastereomeric excess up to 99%. Therefore, we conclude that this bi-enzymatic cascade is capable of producing chiral amines with both high enantiomeric and diastereomeric excess. Further research to perform cascades on a larger scale is recommended.