Pre and post cloning characterization of a beta-1,4-endoglucanase from Bacillus sp

Mol Biol Rep. 2010 Apr;37(4):1717-23. doi: 10.1007/s11033-009-9592-5. Epub 2009 Jun 28.

Abstract

Consistent with its precloning characterization from the cellulolytic Bacillus sp., beta-1,4-endoglucanase purified from the recombinant E. coli exhibited maximum activity at 60 degrees C and pH 7.0. It was highly specific for CMC hydrolysis, with stability up to 70 degrees C and over a pH range of 6.0-8.0. The K(m) and V(max) values for CMCase activity of the enzyme were 4.1 mg/ml and 25 micromole/ml min(-1), respectively. The purified enzyme was a monomer of 65 kDa, as determined by SDS-PAGE. The presence of sucrose and IPTG in fermentation media increased the endoglucanase activity of the recombinant enzyme to 5.2-folds as compared with that of the actual one.

MeSH terms

  • Bacillus / enzymology*
  • Cellulase / genetics*
  • Cellulase / isolation & purification
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Stability
  • Escherichia coli
  • Hydrogen-Ion Concentration
  • Molecular Weight
  • Recombination, Genetic
  • Temperature
  • Transformation, Genetic

Substances

  • Cellulase