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
Descripción
Sumario: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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