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  • Immobilization of pectinase from Leucoagaricus gongylophorus on magnetic particles.

Immobilization of pectinase from Leucoagaricus gongylophorus on magnetic particles.

The Analyst (2012-09-05)
Paulo Roberto Adalberto, Francisco José dos Santos, Camilla Calemi Golfeto, Mônica Rosas Costa Iemma, Dulce Helena Ferreira de Souza, Quezia Bezerra Cass
ABSTRACT

Polygalacturonases (EC 3.2.1.15) hydrolyze the α-1,4-glycosidic linkages in polygalacturonic acid chains. The interest on specific inhibitors of pectinase and the versatility of magnetic support for enzyme immobilization endorsed the preparation of an immobilized enzyme reactor (IMER). This work presents the synthesis of CoFe(2)O(4) amino-derivatives, which was employed as the support for the immobilization of pectinases from Leucoagaricus gongylophorus. Amino-functionalized CoFe(2)O(4) was obtained from glyceryl-derivatized CoFe(2)O(4) and was characterized by infrared spectroscopy and electronic microscopy. The immobilized enzyme maintained the same thermal, chemical and kinetic behaviour of the free enzyme (T(opt) 60 °C; pH(opt) 5.0; K(app)(M) = 0.5 mg min(-1); V(app)(M) ≈ 5.0 μmol min(-1) mL(-1)). The straightforward synthesis of CoFe(2)O(4) derivatives and the efficiency of immobilization offer wide perspectives for the use of the developed new IMER.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Pectolyase from Aspergillus japonicus, suitable for plant cell culture, lyophilized powder
Sigma-Aldrich
Pectinase from Aspergillus niger, BioReagent, suitable for plant cell culture, aqueous glycerol solution, ≥5 units/mg protein (Lowry)
Sigma-Aldrich
Pectolyase from Aspergillus japonicus, lyophilized powder, ≥0.3 units/mg solid
Sigma-Aldrich
Pectinase from Aspergillus niger, powder, slightly beige, >1 U/mg
Sigma-Aldrich
Pectinase from Rhizopus sp., powder, 400-800 units/g solid