title: Identification of (S)-selective transaminases for the asymmetric synthesis of bulky chiral amines.
abstract: The use of transaminases to access pharmaceutically relevant chiral amines is an attractive alternative to transition-metal-catalysed asymmetric chemical synthesis. However, one major challenge is their limited substrate scope. Here we report the creation of highly active and stereoselective transaminases starting from fold class I. The transaminases were developed by extensive protein engineering followed by optimization of the identified motif. The resulting enzymes exhibited up to 8,900-fold higher activity than the starting scaffold and are highly stereoselective (up to >99.9% enantiomeric excess) in the asymmetric synthesis of a set of chiral amines bearing bulky substituents. These enzymes should therefore be suitable for use in the synthesis of a wide array of potential intermediates for pharmaceuticals. We also show that the motif can be engineered into other protein scaffolds with sequence identities as low as 70%, and as such should have a broad impact in the field of biocatalytic synthesis and enzyme engineering.
authors: Department of Biotechnology, Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Str. 4, D-17489 Greifswald, Germany.|Department of Biochemistry, University of Kassel, Heinrich-Plett-Str. 40, D-34132 Kassel, Germany.
authors: Ioannis V
authors: 0000-0001-5811-368X
authors: IV
authors: Pavlidis
authors: Department of Biotechnology, Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Str. 4, D-17489 Greifswald, Germany.
authors: Martin S
authors: MS
authors: Weiß
authors: Department of Biotechnology, Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Str. 4, D-17489 Greifswald, Germany.
authors: Maika
authors: M
authors: Genz
authors: Process Research and Development, Biocatalysis, F. Hoffmann-LaRoche Ltd, Grenzacher Str. 124, 4070 Basel, Switzerland.
authors: Paul
authors: P
authors: Spurr
authors: Process Research and Development, Biocatalysis, F. Hoffmann-LaRoche Ltd, Grenzacher Str. 124, 4070 Basel, Switzerland.
authors: Steven P
authors: SP
authors: Hanlon
authors: Process Research and Development, Biocatalysis, F. Hoffmann-LaRoche Ltd, Grenzacher Str. 124, 4070 Basel, Switzerland.
authors: Beat
authors: B
authors: Wirz
authors: Process Research and Development, Biocatalysis, F. Hoffmann-LaRoche Ltd, Grenzacher Str. 124, 4070 Basel, Switzerland.
authors: Hans
authors: H
authors: Iding
authors: Department of Biotechnology, Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Str. 4, D-17489 Greifswald, Germany.
authors: Uwe T
authors: 0000-0003-0685-2696
authors: UT
authors: Bornscheuer
mesh_terms: D000588:Amines / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D055162:Biocatalysis; D020134:Catalytic Domain; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering; D011809:Quinones / Q000737:chemistry / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity; D000637:Transaminases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000588:Amines; D011809:Quinones; D000637:Transaminases