Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption.

摘要: β-Galactosidase is a hydrolase enzyme that catalyzes the hydrolysis of β-galactosides into monosaccharides; its major application in food industry to reduce content lactose lactic products. The aim this work recover from cell lysate by adsorption onto Streamline-DEAE an expanded bed, avoiding, as much possible, biomass deposition adsorbent matrix. So achieve less debris-matrix interaction, surface was covered with polyvinyl pyrrolidone. showed bind same extent naked and (65 mg β-gal/g matrix) batch mode absence any biomass. kinetics process studied no effect pyrrolidone covering found. optimal conditions for recovery were achieved using made 40% wet weight cells, pyrrolidone-covered matrix/lysate ratio 10% carrying out bed recirculation over 2h 20 mM phosphate buffer pH 7.4. fraction recovered after elution contained 65% initial amount 12.6-fold increased specific activity respect lysate. eluate determined found negligible. remarkable point it possible partially purify feedstock containing unusually high concentration presence weak anion exchangers.