Kinetic Resolution of Racemic Amines to Enantiopure (<i>S</i>)-amines by a Biocatalytic Cascade Employing Amine Dehydrogenase and Alanine Dehydrogenase

Kinetic Resolution of Racemic Amines to Enantiopure (<i>S</i>)-amines by a Biocatalytic Cascade Employing Amine Dehydrogenase and Alanine Dehydrogenase

Amine dehydrogenases (AmDHs) efficiently catalyze the NAD(P)H-dependent asymmetric reductive amination of prochiral carbonyl substrates with high enantioselectivity. AmDH-catalyzed oxidative deamination can also be used for the kinetic resolution of racemic amines to obtain enantiopure amines. In th...

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Main Authors: Mahesh D. Patil, Sanghan Yoon, Hyunwoo Jeon, Taresh P. Khobragade, Sharad Sarak, Amol D. Pagar, Yumi Won, Hyungdon Yun
Format: Article
Language:English
Published: MDPI AG 2019-07-01
Series:Catalysts
Subjects:
amine dehydrogenase
alanine dehydrogenase
chiral amines
kinetic resolution
whole-cell biotransformations
oxidative deamination
Online Access:https://www.mdpi.com/2073-4344/9/7/600
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spelling doaj-f45c841a474346b3ba16f0539d91b8dc2020-11-24T21:28:27ZengMDPI AGCatalysts2073-43442019-07-019760010.3390/catal9070600catal9070600Kinetic Resolution of Racemic Amines to Enantiopure (<i>S</i>)-amines by a Biocatalytic Cascade Employing Amine Dehydrogenase and Alanine DehydrogenaseMahesh D. Patil0Sanghan Yoon1Hyunwoo Jeon2Taresh P. Khobragade3Sharad Sarak4Amol D. Pagar5Yumi Won6Hyungdon Yun7Department of Systems Biotechnology, Konkuk University, Seoul 05029, KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul 05029, KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul 05029, KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul 05029, KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul 05029, KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul 05029, KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul 05029, KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul 05029, KoreaAmine dehydrogenases (AmDHs) efficiently catalyze the NAD(P)H-dependent asymmetric reductive amination of prochiral carbonyl substrates with high enantioselectivity. AmDH-catalyzed oxidative deamination can also be used for the kinetic resolution of racemic amines to obtain enantiopure amines. In the present study, kinetic resolution was carried out using a coupled-enzyme cascade consisting of AmDH and alanine dehydrogenase (AlaDH). AlaDH efficiently catalyzed the conversion of pyruvate to alanine, thus recycling the nicotinamide cofactors and driving the reaction forward. The <i>ee</i> values obtained for the kinetic resolution of 25 and 50 mM <i>rac</i>-&#945;-methylbenzylamine using the purified enzymatic systems were only 54 and 43%, respectively. The use of whole-cells apparently reduced the substrate/product inhibition, and the use of only 30 and 40 mg<sub>DCW</sub>/mL of whole-cells co-expressing AmDH and AlaDH efficiently resolved 100 mM of <i>rac</i>-2-aminoheptane and <i>rac</i>-&#945;-methylbenzylamine into the corresponding enantiopure (<i>S</i>)-amines. Furthermore, the applicability of the reaction protocol demonstrated herein was also successfully tested for the efficient kinetic resolution of wide range of racemic amines.https://www.mdpi.com/2073-4344/9/7/600amine dehydrogenasealanine dehydrogenasechiral amineskinetic resolutionwhole-cell biotransformationsoxidative deamination
collection DOAJ
language English
format Article
sources DOAJ
author Mahesh D. Patil
Sanghan Yoon
Hyunwoo Jeon
Taresh P. Khobragade
Sharad Sarak
Amol D. Pagar
Yumi Won
Hyungdon Yun
spellingShingle Mahesh D. Patil
Sanghan Yoon
Hyunwoo Jeon
Taresh P. Khobragade
Sharad Sarak
Amol D. Pagar
Yumi Won
Hyungdon Yun
Kinetic Resolution of Racemic Amines to Enantiopure (<i>S</i>)-amines by a Biocatalytic Cascade Employing Amine Dehydrogenase and Alanine Dehydrogenase
Catalysts
amine dehydrogenase
alanine dehydrogenase
chiral amines
kinetic resolution
whole-cell biotransformations
oxidative deamination
author_facet Mahesh D. Patil
Sanghan Yoon
Hyunwoo Jeon
Taresh P. Khobragade
Sharad Sarak
Amol D. Pagar
Yumi Won
Hyungdon Yun
author_sort Mahesh D. Patil
title Kinetic Resolution of Racemic Amines to Enantiopure (<i>S</i>)-amines by a Biocatalytic Cascade Employing Amine Dehydrogenase and Alanine Dehydrogenase
title_short Kinetic Resolution of Racemic Amines to Enantiopure (<i>S</i>)-amines by a Biocatalytic Cascade Employing Amine Dehydrogenase and Alanine Dehydrogenase
title_full Kinetic Resolution of Racemic Amines to Enantiopure (<i>S</i>)-amines by a Biocatalytic Cascade Employing Amine Dehydrogenase and Alanine Dehydrogenase
title_fullStr Kinetic Resolution of Racemic Amines to Enantiopure (<i>S</i>)-amines by a Biocatalytic Cascade Employing Amine Dehydrogenase and Alanine Dehydrogenase
title_full_unstemmed Kinetic Resolution of Racemic Amines to Enantiopure (<i>S</i>)-amines by a Biocatalytic Cascade Employing Amine Dehydrogenase and Alanine Dehydrogenase
title_sort kinetic resolution of racemic amines to enantiopure (<i>s</i>)-amines by a biocatalytic cascade employing amine dehydrogenase and alanine dehydrogenase
publisher MDPI AG
series Catalysts
issn 2073-4344
publishDate 2019-07-01
description Amine dehydrogenases (AmDHs) efficiently catalyze the NAD(P)H-dependent asymmetric reductive amination of prochiral carbonyl substrates with high enantioselectivity. AmDH-catalyzed oxidative deamination can also be used for the kinetic resolution of racemic amines to obtain enantiopure amines. In the present study, kinetic resolution was carried out using a coupled-enzyme cascade consisting of AmDH and alanine dehydrogenase (AlaDH). AlaDH efficiently catalyzed the conversion of pyruvate to alanine, thus recycling the nicotinamide cofactors and driving the reaction forward. The <i>ee</i> values obtained for the kinetic resolution of 25 and 50 mM <i>rac</i>-&#945;-methylbenzylamine using the purified enzymatic systems were only 54 and 43%, respectively. The use of whole-cells apparently reduced the substrate/product inhibition, and the use of only 30 and 40 mg<sub>DCW</sub>/mL of whole-cells co-expressing AmDH and AlaDH efficiently resolved 100 mM of <i>rac</i>-2-aminoheptane and <i>rac</i>-&#945;-methylbenzylamine into the corresponding enantiopure (<i>S</i>)-amines. Furthermore, the applicability of the reaction protocol demonstrated herein was also successfully tested for the efficient kinetic resolution of wide range of racemic amines.
topic amine dehydrogenase
alanine dehydrogenase
chiral amines
kinetic resolution
whole-cell biotransformations
oxidative deamination
url https://www.mdpi.com/2073-4344/9/7/600
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