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Methods Enzymol ; 608: 131-149, 2018.
Article in English | MEDLINE | ID: mdl-30173761

ABSTRACT

Synthesis of the chiral amine moiety is a key challenge for synthetic organic chemistry due to its prevalence in many biologically active molecules. Imine reductase and amine oxidase enzymes have enabled the biocatalytic synthesis of a host of chiral amine compounds. In this chapter, procedures for the synthesis of chiral amines using imine reductases (IREDs), the recently discovered IRED homologues reductive aminases, and amine oxidases (AOs) are described. Amine oxidases have been the subject of mutagenesis approaches for improvement of substrate scope. The high-throughput screening method for determining active variants in amine oxidase libraries is illustrated. Finally, in an approach which takes inspiration from nature, many enzymes can be combined with each other in cascade reactions. The incorporation of imine reductase and monoamine oxidase biocatalysts into several cascade reactions, both in vitro and in vivo (where the approach moves toward synthetic biology), is reported.


Subject(s)
Amines/metabolism , Aminohydrolases/metabolism , Bacteria/enzymology , Fungi/enzymology , Monoamine Oxidase/metabolism , Oxidoreductases/metabolism , Protein Engineering/methods , Amines/chemistry , Aminohydrolases/genetics , Aspergillus niger/enzymology , Aspergillus niger/genetics , Aspergillus niger/metabolism , Bacteria/genetics , Bacteria/metabolism , Biocatalysis , Escherichia coli/enzymology , Escherichia coli/genetics , Escherichia coli/metabolism , Fungi/genetics , Fungi/metabolism , Imines/chemistry , Imines/metabolism , Monoamine Oxidase/genetics , Oxidation-Reduction , Oxidoreductases/genetics , Stereoisomerism , Streptomyces/enzymology , Streptomyces/genetics , Streptomyces/metabolism , Synthetic Biology/methods
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