Beta-galactosidase of Penicillium chrysogenum: production, purification, and characterization of the enzyme

Protein Expr Purif. 2001 Feb;21(1):24-9. doi: 10.1006/prep.2000.1344.

Abstract

Intracellular beta-galactosidase from Penicillium chrysogenum NCAIM 00237 was purified by procedures including precipitation with ammonium sulfate, ion-exchange chromatography on DEAE-Sephadex, affinity chromatography, and chromatofocusing. These steps resulted a purification of 66-fold, a yield of about 8%, and a specific activity of 5.84 U mg(-1) protein. Some enzyme characteristics were determined using o-nitrophenyl-beta-d-galactopyranoside as substrate. The pH and temperature optimum of the activity were about 4.0 and 30 degrees C respectively. The K(m) and pI values were 1.81 mM and 4.6. beta-Galactosidase of P. chrysogenum is a multimeric enzyme of about 270 kDa composed of monomers with a molecular mass of 66 kDa.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, Affinity / methods
  • Chromatography, Ion Exchange / methods
  • Electrophoresis, Polyacrylamide Gel
  • Kinetics
  • Molecular Weight
  • Penicillium chrysogenum / enzymology*
  • Thermodynamics
  • Ultrafiltration
  • beta-Galactosidase / chemistry
  • beta-Galactosidase / isolation & purification*
  • beta-Galactosidase / metabolism*

Substances

  • beta-Galactosidase