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A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria

Nitrile hydrolyzing enzymes catalyze the hydration of nitrile compounds to corresponding amides and acids. Bacteria, isolated from soil samples were screened for nitrile hydrolyzing enzymes by simple dye based 96 well plate and nesslerization method. Bromothymol blue was used as an indicator for the...

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Detalles Bibliográficos
Autores principales: Sahu, Ruchi, Meghavarnam, Anil Kumar, Janakiraman, Savitha
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6881679/
https://www.ncbi.nlm.nih.gov/pubmed/31799145
http://dx.doi.org/10.1016/j.btre.2019.e00396
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author Sahu, Ruchi
Meghavarnam, Anil Kumar
Janakiraman, Savitha
author_facet Sahu, Ruchi
Meghavarnam, Anil Kumar
Janakiraman, Savitha
author_sort Sahu, Ruchi
collection PubMed
description Nitrile hydrolyzing enzymes catalyze the hydration of nitrile compounds to corresponding amides and acids. Bacteria, isolated from soil samples were screened for nitrile hydrolyzing enzymes by simple dye based 96 well plate and nesslerization method. Bromothymol blue was used as an indicator for the detection of amides and acids based on colour change of the indicator dye from blue to dark green or yellow. The screening assay also differentiates between nitrile hydratase (NHase) and nitrilase producing bacteria. Among the 108 bacterial strains screened for enzyme activity, six strains were positive for NHase activity and eleven strains were positive for nitrilase activity based on their ability to degrade acrylonitrile into products. The strain showing maximum NHase activity in quantitative assay was identified as Rhodococcus rhodochrous. The modified method developed by us would be useful for rapid screening of nitrile degrading bacteria potent for acrylamide/acrylic acid production when acrylonitrile is supplied as substrate.
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spelling pubmed-68816792019-12-03 A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria Sahu, Ruchi Meghavarnam, Anil Kumar Janakiraman, Savitha Biotechnol Rep (Amst) Research Article Nitrile hydrolyzing enzymes catalyze the hydration of nitrile compounds to corresponding amides and acids. Bacteria, isolated from soil samples were screened for nitrile hydrolyzing enzymes by simple dye based 96 well plate and nesslerization method. Bromothymol blue was used as an indicator for the detection of amides and acids based on colour change of the indicator dye from blue to dark green or yellow. The screening assay also differentiates between nitrile hydratase (NHase) and nitrilase producing bacteria. Among the 108 bacterial strains screened for enzyme activity, six strains were positive for NHase activity and eleven strains were positive for nitrilase activity based on their ability to degrade acrylonitrile into products. The strain showing maximum NHase activity in quantitative assay was identified as Rhodococcus rhodochrous. The modified method developed by us would be useful for rapid screening of nitrile degrading bacteria potent for acrylamide/acrylic acid production when acrylonitrile is supplied as substrate. Elsevier 2019-11-15 /pmc/articles/PMC6881679/ /pubmed/31799145 http://dx.doi.org/10.1016/j.btre.2019.e00396 Text en © 2019 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Article
Sahu, Ruchi
Meghavarnam, Anil Kumar
Janakiraman, Savitha
A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria
title A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria
title_full A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria
title_fullStr A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria
title_full_unstemmed A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria
title_short A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria
title_sort simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6881679/
https://www.ncbi.nlm.nih.gov/pubmed/31799145
http://dx.doi.org/10.1016/j.btre.2019.e00396
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