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

Extremophiles. 2010 Mar;14(2):185-92. doi: 10.1007/s00792-009-0297-4.

Abstract

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 x 10(-9) s (-1) and a K (m) of 206 microM 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 degrees C and the enzyme had a half life at 60 degrees C of 20.8 h.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacillus / enzymology*
  • Bacillus / genetics*
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers / genetics
  • DNA, Bacterial / genetics
  • Escherichia coli / genetics
  • Genes, Bacterial
  • Hydrogen-Ion Concentration
  • Kinetics
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Structure, Tertiary
  • Purine-Nucleoside Phosphorylase / genetics*
  • Purine-Nucleoside Phosphorylase / isolation & purification*
  • Purine-Nucleoside Phosphorylase / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Structural Homology, Protein
  • Thermodynamics

Substances

  • DNA Primers
  • DNA, Bacterial
  • Recombinant Proteins
  • Purine-Nucleoside Phosphorylase

Associated data

  • GENBANK/GQ390428