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The identification and use of robust transaminases from a domestic drain metagenome

Transaminases remain one of the most promising biocatalysts for use in chiral amine synthesis, however their industrial implementation has been hampered by their general instability towards, for example, high amine donor concentrations and organic solvent content. Herein we describe the identificati...

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Autores principales: Leipold, Leona, Dobrijevic, Dragana, Jeffries, Jack W. E., Bawn, Maria, Moody, Thomas S., Ward, John M., Hailes, Helen C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Royal Society of Chemistry 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6394892/
https://www.ncbi.nlm.nih.gov/pubmed/30930686
http://dx.doi.org/10.1039/c8gc02986e
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author Leipold, Leona
Dobrijevic, Dragana
Jeffries, Jack W. E.
Bawn, Maria
Moody, Thomas S.
Ward, John M.
Hailes, Helen C.
author_facet Leipold, Leona
Dobrijevic, Dragana
Jeffries, Jack W. E.
Bawn, Maria
Moody, Thomas S.
Ward, John M.
Hailes, Helen C.
author_sort Leipold, Leona
collection PubMed
description Transaminases remain one of the most promising biocatalysts for use in chiral amine synthesis, however their industrial implementation has been hampered by their general instability towards, for example, high amine donor concentrations and organic solvent content. Herein we describe the identification, cloning and screening of 29 novel transaminases from a household drain metagenome. The most promising enzymes were fully characterised and the effects of pH, temperature, amine donor concentration and co-solvent determined. Several enzymes demonstrated good substrate tolerance as well as an unprecedented robustness for a wild-type transaminase. One enzyme in particular readily accepted IPA as an amine donor giving the same conversion with 2–50 equivalents, as well as being tolerant to a number of co-solvents, and operational in up to 50% DMSO – a characteristic as yet unobserved in a wild-type transaminase. This work highlights the value of using metagenomics for biocatalyst discovery from niche environments, and here has led to the identification of one of the most robust native transaminases described to date, with respect to IPA and DMSO tolerance.
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spelling pubmed-63948922019-03-29 The identification and use of robust transaminases from a domestic drain metagenome Leipold, Leona Dobrijevic, Dragana Jeffries, Jack W. E. Bawn, Maria Moody, Thomas S. Ward, John M. Hailes, Helen C. Green Chem Chemistry Transaminases remain one of the most promising biocatalysts for use in chiral amine synthesis, however their industrial implementation has been hampered by their general instability towards, for example, high amine donor concentrations and organic solvent content. Herein we describe the identification, cloning and screening of 29 novel transaminases from a household drain metagenome. The most promising enzymes were fully characterised and the effects of pH, temperature, amine donor concentration and co-solvent determined. Several enzymes demonstrated good substrate tolerance as well as an unprecedented robustness for a wild-type transaminase. One enzyme in particular readily accepted IPA as an amine donor giving the same conversion with 2–50 equivalents, as well as being tolerant to a number of co-solvents, and operational in up to 50% DMSO – a characteristic as yet unobserved in a wild-type transaminase. This work highlights the value of using metagenomics for biocatalyst discovery from niche environments, and here has led to the identification of one of the most robust native transaminases described to date, with respect to IPA and DMSO tolerance. Royal Society of Chemistry 2019-01-07 2018-11-15 /pmc/articles/PMC6394892/ /pubmed/30930686 http://dx.doi.org/10.1039/c8gc02986e Text en This journal is © The Royal Society of Chemistry 2019 http://creativecommons.org/licenses/by/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence (CC BY 3.0)
spellingShingle Chemistry
Leipold, Leona
Dobrijevic, Dragana
Jeffries, Jack W. E.
Bawn, Maria
Moody, Thomas S.
Ward, John M.
Hailes, Helen C.
The identification and use of robust transaminases from a domestic drain metagenome
title The identification and use of robust transaminases from a domestic drain metagenome
title_full The identification and use of robust transaminases from a domestic drain metagenome
title_fullStr The identification and use of robust transaminases from a domestic drain metagenome
title_full_unstemmed The identification and use of robust transaminases from a domestic drain metagenome
title_short The identification and use of robust transaminases from a domestic drain metagenome
title_sort identification and use of robust transaminases from a domestic drain metagenome
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6394892/
https://www.ncbi.nlm.nih.gov/pubmed/30930686
http://dx.doi.org/10.1039/c8gc02986e
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