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A New Thermophilic Nitrilase from an Antarctic Hyperthermophilic Microorganism

Several environmental samples from Antarctica were collected and enriched to search for microorganisms with nitrilase activity. A new thermostable nitrilase from a novel hyperthermophilic archaea Pyrococcus sp. M24D13 was purified and characterized. The activity of this enzyme increased as the tempe...

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Autores principales: Dennett, Geraldine V., Blamey, Jenny M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4770253/
https://www.ncbi.nlm.nih.gov/pubmed/26973832
http://dx.doi.org/10.3389/fbioe.2016.00005
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author Dennett, Geraldine V.
Blamey, Jenny M.
author_facet Dennett, Geraldine V.
Blamey, Jenny M.
author_sort Dennett, Geraldine V.
collection PubMed
description Several environmental samples from Antarctica were collected and enriched to search for microorganisms with nitrilase activity. A new thermostable nitrilase from a novel hyperthermophilic archaea Pyrococcus sp. M24D13 was purified and characterized. The activity of this enzyme increased as the temperatures rise from 70 up to 85°C. Its optimal activity occurred at 85°C and pH 7.5. This new enzyme shows a remarkable resistance to thermal inactivation retaining more than 50% of its activity even after 8 h of incubation at 85°C. In addition, this nitrilase is highly versatile demonstrating activity toward different substrates, such as benzonitrile (60 mM, aromatic nitrile) and butyronitrile (60 mM, aliphatic nitrile), with a specific activity of 3286.7 U mg(−1) of protein and 4008.2 U mg(−1) of protein, respectively. Moreover the enzyme NitM24D13 also presents cyanidase activity. The apparent Michaelis–Menten constant (K(m)) and V(máx) of this Nitrilase for benzonitrile were 0.3 mM and 333.3 μM min(−1), respectively, and the specificity constant (k(cat)/K(m)) for benzonitrile was 2.05 × 10(5) s(−1) M(−1).
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spelling pubmed-47702532016-03-11 A New Thermophilic Nitrilase from an Antarctic Hyperthermophilic Microorganism Dennett, Geraldine V. Blamey, Jenny M. Front Bioeng Biotechnol Bioengineering and Biotechnology Several environmental samples from Antarctica were collected and enriched to search for microorganisms with nitrilase activity. A new thermostable nitrilase from a novel hyperthermophilic archaea Pyrococcus sp. M24D13 was purified and characterized. The activity of this enzyme increased as the temperatures rise from 70 up to 85°C. Its optimal activity occurred at 85°C and pH 7.5. This new enzyme shows a remarkable resistance to thermal inactivation retaining more than 50% of its activity even after 8 h of incubation at 85°C. In addition, this nitrilase is highly versatile demonstrating activity toward different substrates, such as benzonitrile (60 mM, aromatic nitrile) and butyronitrile (60 mM, aliphatic nitrile), with a specific activity of 3286.7 U mg(−1) of protein and 4008.2 U mg(−1) of protein, respectively. Moreover the enzyme NitM24D13 also presents cyanidase activity. The apparent Michaelis–Menten constant (K(m)) and V(máx) of this Nitrilase for benzonitrile were 0.3 mM and 333.3 μM min(−1), respectively, and the specificity constant (k(cat)/K(m)) for benzonitrile was 2.05 × 10(5) s(−1) M(−1). Frontiers Media S.A. 2016-02-29 /pmc/articles/PMC4770253/ /pubmed/26973832 http://dx.doi.org/10.3389/fbioe.2016.00005 Text en Copyright © 2016 Dennett and Blamey. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Dennett, Geraldine V.
Blamey, Jenny M.
A New Thermophilic Nitrilase from an Antarctic Hyperthermophilic Microorganism
title A New Thermophilic Nitrilase from an Antarctic Hyperthermophilic Microorganism
title_full A New Thermophilic Nitrilase from an Antarctic Hyperthermophilic Microorganism
title_fullStr A New Thermophilic Nitrilase from an Antarctic Hyperthermophilic Microorganism
title_full_unstemmed A New Thermophilic Nitrilase from an Antarctic Hyperthermophilic Microorganism
title_short A New Thermophilic Nitrilase from an Antarctic Hyperthermophilic Microorganism
title_sort new thermophilic nitrilase from an antarctic hyperthermophilic microorganism
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4770253/
https://www.ncbi.nlm.nih.gov/pubmed/26973832
http://dx.doi.org/10.3389/fbioe.2016.00005
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