Amine Transaminase Engineering for Spatially Bulky Substrate Acceptance.
Amine transaminase (ATA) catalyzing stereoselective amination of prochiral ketones is an attractive alternative to transition metal catalysis. As wild-type ATAs do not accept sterically hindered ketones, efforts to widen the substrate scope to more challenging targets are of general interest. We recently designed ATAs to accept aromatic and thus planar bulky amines, with a sequence-based motif that supports the identification of novel enzymes. However, these variants were not active against 2,2-dimethyl-1-phenyl-propan-1-one, which carries a bulky tert-butyl substituent adjacent to the carbonyl function. Here, we report a solution for this type of substrate. The evolved ATAs perform asymmetric synthesis of the respective R amine with high conversions by using either alanine or isopropylamine as amine donor.
Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17489, Greifswald, Germany.
Martin S
MS
Weiß
Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17489, Greifswald, Germany.|Group of Biotechnology, Department of Biochemistry, University of Kassel, Heinrich-Plett-Strasse 40, 34132, Kassel, Germany.
Ioannis V
0000-0001-5811-368X
IV
Pavlidis
Process Chemistry and Catalysis, Biocatalysis, F. Hoffmann-La Roche Ltd., Grenzacher Strasse 124, 4070, Basel, Switzerland.
Paul
P
Spurr
Process Chemistry and Catalysis, Biocatalysis, F. Hoffmann-La Roche Ltd., Grenzacher Strasse 124, 4070, Basel, Switzerland.
Steven P
SP
Hanlon
Process Chemistry and Catalysis, Biocatalysis, F. Hoffmann-La Roche Ltd., Grenzacher Strasse 124, 4070, Basel, Switzerland.
Beat
B
Wirz
Process Chemistry and Catalysis, Biocatalysis, F. Hoffmann-La Roche Ltd., Grenzacher Strasse 124, 4070, Basel, Switzerland.
Hans
H
Iding
Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17489, Greifswald, Germany.
Uwe T
0000-0003-0685-2696
UT
Bornscheuer
D000586:Amination; D000588:Amines; D019943:Amino Acid Substitution; D055162:Biocatalysis; D003198:Computer Simulation; D019020:Directed Molecular Evolution; D015202:Protein Engineering / Q000379:methods*; D013379:Substrate Specificity; D000637:Transaminases / Q000235:genetics*
D000588:Amines; D000637:Transaminases
amine transaminase; asymmetric synthesis; enzyme catalysis; protein engineering