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

doi:10.1021/acscatal.6b02380

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

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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Abstract

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 language	English
Pages (from-to)	7749
Journal	ACS Catalysis
Volume	6
Issue number	11
Early online date	04 Oct 2016
DOIs	https://doi.org/10.1021/acscatal.6b02380
Publication status	Early online date - 04 Oct 2016

Bibliographical note

This is a interdisciplinary research in collaboration with experimentalists from Almac and Queens.

The work was funded by the Invest NI project RD11181114 (2014-2019).

ACS catalysis is a peer-reviewed journal with an Impact Factor of 13.1

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10.1021/acscatal.6b02380Licence: Unspecified

Rational design of a (S)-selective-Transaminase for asymmetric synthesis of (1S)-1-(1,1'-biphenyl 2-yl)ethanamine
This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Catalysis, copyright © 2016 American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see: http://pubs.acs.org/doi/abs/10.1021/acscatal.6b02380
Accepted author manuscript, 854 KBLicence: Unspecified

Christopher C R Allen
- C.Allenqub.acuk
Person: Academic
Meilan Huang
- m.huangqub.acuk
- School of Chemistry and Chemical Engineering - Senior Lecturer
- Theoretical and Applied Catalysis
Person: Academic
Tim Skvortsov
- t.skvortsovqub.acuk
- School of Pharmacy - Lecturer
- Material and Advanced Technologies for Healthcare
Person: Academic

Cite this

Dourado, D. F. A. R., Pohle, S., Carvalho, A. T. P., Dheeman, D. S., Caswell, J. M., Skvortsov, T., Miskelly, I., Brown, R. T., Quinn, D. J., Allen, C. C. R., Kulakov, L., Huang, M., & Moody, T. S. (2016). Rational design of a (S)-selective-Transaminase for asymmetric synthesis of (1S)-1-(1,1'-biphenyl 2-yl)ethanamine. ACS Catalysis, 6(11), 7749. Advance online publication. https://doi.org/10.1021/acscatal.6b02380

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

abstract = "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%). ",

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

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AU - Pohle, Stefan

AU - Carvalho, Alexandra T. P.

AU - Dheeman, Dharmendra S.

AU - Caswell, Jill M.

AU - Skvortsov, Timofey

AU - Miskelly, Iain

AU - Brown, Rodney T.

AU - Quinn, Derek J.

AU - Allen, Christopher C. R.

AU - Kulakov, Leonid

AU - Huang, Meilan

AU - Moody, Thomas S.

N1 - This is a interdisciplinary research in collaboration with experimentalists from Almac and Queens. The work was funded by the Invest NI project RD11181114 (2014-2019). ACS catalysis is a peer-reviewed journal with an Impact Factor of 13.1

PY - 2016/10/4

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N2 - 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%).

AB - 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%).

U2 - 10.1021/acscatal.6b02380

DO - 10.1021/acscatal.6b02380

M3 - Article

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

Abstract

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Christopher C R Allen

Meilan Huang

Tim Skvortsov

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