Rational design of a (S)-selective-Transaminase for asymmetric synthesis of (1S)-1-(1,1'-biphenyl 2-yl)ethanamine

Daniel F. A. R. Dourado, Stefan Pohle, Alexandra T. P. Carvalho, Dharmendra S. Dheeman, Jill M. Caswell, Timofey Skvortsov, Iain Miskelly, Rodney T. Brown, Derek J. Quinn, Christopher C. R. Allen, Leonid Kulakov, Meilan Huang, Thomas S. Moody

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35 Citations (Scopus)
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Amine transaminases offer an environmentally sustainable synthesis route for the production ofpure chiral amines. However, their catalytic efficiency towards bulky ketone substrates isgreatly limited by steric hindrance and therefore presents a great challenge for industrialsynthetic applications. Hereby we report an example of rational transaminase enzyme design tohelp alleviate these challenges. Starting from the Vibrio fluvialis amine transaminase that has nodetectable catalytic activity towards the bulky aromatic ketone 2-acetylbiphenyl, we employed arational design strategy combining in silico and in vitro studies to engineer the transaminaseenzyme with a minimal number of mutations, achieving an high catalytic activity and highenantioselectivity. We found that by introducing two mutations W57G/R415A detectableenzyme activity was achieved. The rationally designed best variant,W57F/R88H/V153S/K163F/I259M/R415A/V422A, showed an improvement in reaction rateby > 1716-fold towards the bulky ketone under study, producing the corresponding enantiomericpure (S)-amine (ee value of > 99%). 
Original languageEnglish
Pages (from-to)7749
JournalACS Catalysis
Issue number11
Early online date04 Oct 2016
Publication statusEarly online date - 04 Oct 2016


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