Immobilization of β-galactosidase from Kluyveromyces lactis on silica and agarose: comparison of different methods

摘要: Abstract The covalent immobilization of β -galactosidase from Kluyveromyces lactis ( -gal) on to two different porous carriers, CPC-silica and agarose, is reported. was silanizated activated with glutaraldehyde. activation agarose via a cyanylating agent (CDAP) optimized. Gel-bound protein gel-bound activity were both measured directly, allowing the determination apparent specific activities (S.A.). Higher amounts -gal immobilized (maximum capacity, 23 mg ml −1 packed support) than CDAP-activated agarose. For lower enzyme loading assayed (12.6 support), 100% but only 34% its expressed. This inactivation during confirmed by S.A. values (22–29 EU for CPC-derivatives 80 soluble -gal). K app (3.4 mM) CDAP-derivative ONPG as substrate higher M value (2 mM). When increased five-fold, four-fold, 13 mM. V remarkably max -galactosidase. CDAP-derivatives showed better thermal stabilities neither them enhanced stability enzyme. stored at 4°C, derivatives remained stable least 2 months. Both displayed high percentages lactose conversion (90%) in bed mini-reactors. Glucose production 3.3-fold CPC-derivative CDAP-derivative, consequence flow rates achieved.