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Recombinant l‐Amino Acid Oxidase with Broad Substrate Spectrum for Co‐substrate Recycling in (S)‐Selective Transaminase‐Catalyzed Kinetic Resolutions

Chiral and enantiopure amines can be produced by enantioselective transaminases via kinetic resolution of amine racemates. This transamination reaction requires stoichiometric amounts of co‐substrate. A dual‐enzyme recycling system overcomes this limitation: l‐amino acid oxidases (LAAO) recycle the...

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Autores principales: Heinks, Tobias, Paulus, Jannik, Koopmeiners, Simon, Beuel, Tobias, Sewald, Norbert, Höhne, Matthias, Bornscheuer, Uwe T., Fischer von Mollard, Gabriele
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9543090/
https://www.ncbi.nlm.nih.gov/pubmed/35713203
http://dx.doi.org/10.1002/cbic.202200329
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author Heinks, Tobias
Paulus, Jannik
Koopmeiners, Simon
Beuel, Tobias
Sewald, Norbert
Höhne, Matthias
Bornscheuer, Uwe T.
Fischer von Mollard, Gabriele
author_facet Heinks, Tobias
Paulus, Jannik
Koopmeiners, Simon
Beuel, Tobias
Sewald, Norbert
Höhne, Matthias
Bornscheuer, Uwe T.
Fischer von Mollard, Gabriele
author_sort Heinks, Tobias
collection PubMed
description Chiral and enantiopure amines can be produced by enantioselective transaminases via kinetic resolution of amine racemates. This transamination reaction requires stoichiometric amounts of co‐substrate. A dual‐enzyme recycling system overcomes this limitation: l‐amino acid oxidases (LAAO) recycle the accumulating co‐product of (S)‐selective transaminases in the kinetic resolution of racemic amines to produce pure (R)‐amines. However, availability of suitable LAAOs is limited. Here we use the heterologously produced, highly active fungal hcLAAO4 with broad substrate spectrum. H(2)O(2) as byproduct of hcLAAO4 is detoxified by a catalase. The final system allows using sub‐stoichiometric amounts of 1 mol% of the transaminase co‐substrate as well as the initial application of l‐amino acids instead of α‐keto acids. With an optimized protocol, the synthetic potential of this kinetic resolution cascade was proven at the preparative scale (>90 mg) by the synthesis of highly enantiomerically pure (R)‐methylbenzylamine (>99 %ee) at complete conversion (50 %).
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spelling pubmed-95430902022-10-14 Recombinant l‐Amino Acid Oxidase with Broad Substrate Spectrum for Co‐substrate Recycling in (S)‐Selective Transaminase‐Catalyzed Kinetic Resolutions Heinks, Tobias Paulus, Jannik Koopmeiners, Simon Beuel, Tobias Sewald, Norbert Höhne, Matthias Bornscheuer, Uwe T. Fischer von Mollard, Gabriele Chembiochem Research Articles Chiral and enantiopure amines can be produced by enantioselective transaminases via kinetic resolution of amine racemates. This transamination reaction requires stoichiometric amounts of co‐substrate. A dual‐enzyme recycling system overcomes this limitation: l‐amino acid oxidases (LAAO) recycle the accumulating co‐product of (S)‐selective transaminases in the kinetic resolution of racemic amines to produce pure (R)‐amines. However, availability of suitable LAAOs is limited. Here we use the heterologously produced, highly active fungal hcLAAO4 with broad substrate spectrum. H(2)O(2) as byproduct of hcLAAO4 is detoxified by a catalase. The final system allows using sub‐stoichiometric amounts of 1 mol% of the transaminase co‐substrate as well as the initial application of l‐amino acids instead of α‐keto acids. With an optimized protocol, the synthetic potential of this kinetic resolution cascade was proven at the preparative scale (>90 mg) by the synthesis of highly enantiomerically pure (R)‐methylbenzylamine (>99 %ee) at complete conversion (50 %). John Wiley and Sons Inc. 2022-07-05 2022-08-17 /pmc/articles/PMC9543090/ /pubmed/35713203 http://dx.doi.org/10.1002/cbic.202200329 Text en © 2022 The Authors. ChemBioChem published by Wiley-VCH GmbH https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Research Articles
Heinks, Tobias
Paulus, Jannik
Koopmeiners, Simon
Beuel, Tobias
Sewald, Norbert
Höhne, Matthias
Bornscheuer, Uwe T.
Fischer von Mollard, Gabriele
Recombinant l‐Amino Acid Oxidase with Broad Substrate Spectrum for Co‐substrate Recycling in (S)‐Selective Transaminase‐Catalyzed Kinetic Resolutions
title Recombinant l‐Amino Acid Oxidase with Broad Substrate Spectrum for Co‐substrate Recycling in (S)‐Selective Transaminase‐Catalyzed Kinetic Resolutions
title_full Recombinant l‐Amino Acid Oxidase with Broad Substrate Spectrum for Co‐substrate Recycling in (S)‐Selective Transaminase‐Catalyzed Kinetic Resolutions
title_fullStr Recombinant l‐Amino Acid Oxidase with Broad Substrate Spectrum for Co‐substrate Recycling in (S)‐Selective Transaminase‐Catalyzed Kinetic Resolutions
title_full_unstemmed Recombinant l‐Amino Acid Oxidase with Broad Substrate Spectrum for Co‐substrate Recycling in (S)‐Selective Transaminase‐Catalyzed Kinetic Resolutions
title_short Recombinant l‐Amino Acid Oxidase with Broad Substrate Spectrum for Co‐substrate Recycling in (S)‐Selective Transaminase‐Catalyzed Kinetic Resolutions
title_sort recombinant l‐amino acid oxidase with broad substrate spectrum for co‐substrate recycling in (s)‐selective transaminase‐catalyzed kinetic resolutions
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9543090/
https://www.ncbi.nlm.nih.gov/pubmed/35713203
http://dx.doi.org/10.1002/cbic.202200329
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