Generation of Oxidoreductases with Dual Alcohol Dehydrogenase and Amine Dehydrogenase Activity
| Autor: | Vasilis Tseliou, Don Schilder, Marcelo F. Masman, Francesco G. Mutti, Tanja Knaus |
|---|---|
| Přispěvatelé: | Biocatalysis (HIMS, FNWI) |
| Rok vydání: | 2021 |
| Předmět: |
biocatalysis
Alcohol 010402 general chemistry Formate dehydrogenase 01 natural sciences Reductive amination alcohol amination Catalysis chemistry.chemical_compound amine dehydrogenases Biocatalysis | Hot Paper Amines Alcohol dehydrogenase Amination alcohol dehydrogenases biology Full Paper enzyme promiscuity 010405 organic chemistry Organic Chemistry Amine dehydrogenase General Chemistry Full Papers Combinatorial chemistry 0104 chemical sciences chemistry Benzyl alcohol biology.protein Enzyme promiscuity Oxidoreductases Amine dehydrogenase activity |
| Zdroj: | Chemistry – A European Journal Chemistry-A European Journal, 27(10), 3315-3325. Wiley-VCH Verlag Chemistry (Weinheim an Der Bergstrasse, Germany) |
| ISSN: | 0947-6539 |
| DOI: | 10.1002/chem.202003140 |
| Popis: | The l‐lysine‐ϵ‐dehydrogenase (LysEDH) from Geobacillus stearothermophilus naturally catalyzes the oxidative deamination of the ϵ‐amino group of l‐lysine. We previously engineered this enzyme to create amine dehydrogenase (AmDH) variants that possess a new hydrophobic cavity in their active site such that aromatic ketones can bind and be converted into α‐chiral amines with excellent enantioselectivity. We also recently observed that LysEDH was capable of reducing aromatic aldehydes into primary alcohols. Herein, we harnessed the promiscuous alcohol dehydrogenase (ADH) activity of LysEDH to create new variants that exhibited enhanced catalytic activity for the reduction of substituted benzaldehydes and arylaliphatic aldehydes to primary alcohols. Notably, these novel engineered dehydrogenases also catalyzed the reductive amination of a variety of aldehydes and ketones with excellent enantioselectivity, thus exhibiting a dual AmDH/ADH activity. We envisioned that the catalytic bi‐functionality of these enzymes could be applied for the direct conversion of alcohols into amines. As a proof‐of‐principle, we performed an unprecedented one‐pot “hydrogen‐borrowing” cascade to convert benzyl alcohol to benzylamine using a single enzyme. Conducting the same biocatalytic cascade in the presence of cofactor recycling enzymes (i.e., NADH‐oxidase and formate dehydrogenase) increased the reaction yields. In summary, this work provides the first examples of enzymes showing “alcohol aminase” activity. Lend me your hydrogen: Starting from an ϵ‐deaminating l‐lysine dehydrogenase, we created new variants that perform the reduction of aldehydes to alcohols and the reductive amination of aldehydes and ketones to amines. The reductive amination of ketones proceeded with excellent stereoselectivity. This dual AmDH/ADH activity was applied for the one‐enzyme hydrogen‐borrowing amination of benzylic alcohol, thus providing the first example of an “alcohol aminase” activity. |
| Databáze: | OpenAIRE |
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