Cloning, purification and characterisation of a recombinant purine nucleoside phosphorylase from Bacillus halodurans Alk36

A purine nucleoside phosphorylase from the alkaliphile Bacillus halodurans Alk36 was cloned and overexpressed in Escherichia coli. The enzyme was purified fivefold by membrane filtration and ion exchange. The purified enzyme had a V (max) of 2.03 × 10(−9) s (−1) and a K (m) of 206 μM on guanosine. T...

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Detalles Bibliográficos
Autores principales: Visser, Daniel F., Hennessy, Fritha, Rashamuse, Konanani, Louw, Maureen E., Brady, Dean
Formato: Texto
Lenguaje:English
Publicado: Springer Japan 2010
Materias:
Original Paper
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2832885/
https://www.ncbi.nlm.nih.gov/pubmed/20063024
http://dx.doi.org/10.1007/s00792-009-0297-4
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author Visser, Daniel F.
Hennessy, Fritha
Rashamuse, Konanani
Louw, Maureen E.
Brady, Dean
author_facet Visser, Daniel F.
Hennessy, Fritha
Rashamuse, Konanani
Louw, Maureen E.
Brady, Dean
author_sort Visser, Daniel F.
collection PubMed
description A purine nucleoside phosphorylase from the alkaliphile Bacillus halodurans Alk36 was cloned and overexpressed in Escherichia coli. The enzyme was purified fivefold by membrane filtration and ion exchange. The purified enzyme had a V (max) of 2.03 × 10(−9) s (−1) and a K (m) of 206 μM on guanosine. The optimal pH range was between 5.7 and 8.4 with a maximum at pH 7.0. The optimal temperature for activity was 70°C and the enzyme had a half life at 60°C of 20.8 h.
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spelling pubmed-28328852010-03-15 Cloning, purification and characterisation of a recombinant purine nucleoside phosphorylase from Bacillus halodurans Alk36 Visser, Daniel F. Hennessy, Fritha Rashamuse, Konanani Louw, Maureen E. Brady, Dean Extremophiles Original Paper A purine nucleoside phosphorylase from the alkaliphile Bacillus halodurans Alk36 was cloned and overexpressed in Escherichia coli. The enzyme was purified fivefold by membrane filtration and ion exchange. The purified enzyme had a V (max) of 2.03 × 10(−9) s (−1) and a K (m) of 206 μM on guanosine. The optimal pH range was between 5.7 and 8.4 with a maximum at pH 7.0. The optimal temperature for activity was 70°C and the enzyme had a half life at 60°C of 20.8 h. Springer Japan 2010-01-09 2010 /pmc/articles/PMC2832885/ /pubmed/20063024 http://dx.doi.org/10.1007/s00792-009-0297-4 Text en © The Author(s) 2010 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.
spellingShingle Original Paper
Visser, Daniel F.
Hennessy, Fritha
Rashamuse, Konanani
Louw, Maureen E.
Brady, Dean
Cloning, purification and characterisation of a recombinant purine nucleoside phosphorylase from Bacillus halodurans Alk36
title Cloning, purification and characterisation of a recombinant purine nucleoside phosphorylase from Bacillus halodurans Alk36
title_full Cloning, purification and characterisation of a recombinant purine nucleoside phosphorylase from Bacillus halodurans Alk36
title_fullStr Cloning, purification and characterisation of a recombinant purine nucleoside phosphorylase from Bacillus halodurans Alk36
title_full_unstemmed Cloning, purification and characterisation of a recombinant purine nucleoside phosphorylase from Bacillus halodurans Alk36
title_short Cloning, purification and characterisation of a recombinant purine nucleoside phosphorylase from Bacillus halodurans Alk36
title_sort cloning, purification and characterisation of a recombinant purine nucleoside phosphorylase from bacillus halodurans alk36
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2832885/
https://www.ncbi.nlm.nih.gov/pubmed/20063024
http://dx.doi.org/10.1007/s00792-009-0297-4
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