Photometric Characterization of the Reductive Amination Scope of the Imine Reductases fromStreptomyces tsukubaensisandStreptomyces ipomoeae
Autor: | Matthias Höhne, Lukas Krautschick, Philipp Matzel |
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Rok vydání: | 2017 |
Předmět: |
Streptomyces tsukubaensis
Stereochemistry Imine 010402 general chemistry 01 natural sciences Biochemistry Reductive amination Pyrrolidine Photometry chemistry.chemical_compound Nucleophile Organic chemistry Molecular Biology Amination Enzyme Assays biology 010405 organic chemistry Organic Chemistry Substrate (chemistry) Streptomyces ipomoeae biology.organism_classification Streptomyces 0104 chemical sciences chemistry Molecular Medicine Imines Oxidoreductases NADP |
Zdroj: | ChemBioChem. 18:2022-2027 |
ISSN: | 1439-4227 |
DOI: | 10.1002/cbic.201700257 |
Popis: | Imine reductases (IREDs) have emerged as promising enzymes for the asymmetric synthesis of secondary and tertiary amines starting from carbonyl substrates. Screening the substrate specificity of the reductive amination reaction is usually performed by time-consuming GC analytics. We found two highly active IREDs in our enzyme collection, IR-20 from Streptomyces tsukubaensis and IR-Sip from Streptomyces ipomoeae, that allowed a comprehensive substrate screening with a photometric NADPH assay. We screened 39 carbonyl substrates combined with 17 amines as nucleophiles. Activity data from 663 combinations provided a clear picture about substrate specificity and capabilities in the reductive amination of these enzymes. Besides aliphatic aldehydes, the IREDs accepted various cyclic (C4 -C8 ) and acyclic ketones, preferentially with methylamine. IR-Sip also accepted a range of primary and secondary amines as nucleophiles. In biocatalytic reactions, IR-Sip converted (R)-3-methylcyclohexanone with dimethylamine or pyrrolidine with high diastereoselectivity (>94-96 % de). The nucleophile acceptor spectrum depended on the carbonyl substrate employed. The conversion of well-accepted substrates could also be detected if crude lysates were employed as the enzyme source. |
Databáze: | OpenAIRE |
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