Simple eco-friendly .-galactosidase immobilization on functionalized magnetic particles for lactose hydrolysis

Kalim Belhacene1, Elena Florentina Grosu2, Alexandra Cristina Blaga2, Pascal Dhulster1, Mariana Pinteala3, Renato Froidevaux1

1 Charles VIOLLETTE Institute Equipe Laboratoire des Proc d s Biologiques, G nie Enzymatique et Microbien (ProBioGEM) EA 1026, Universit Lille 1 Sciences et Technologies, Villeneuve d Ascq, France
2 Gheorghe Asachi Technical University of Iasi, Faculty of Chemical Engineering and Environmental Protection, Department of Organic, Biochemical and Food Engineering, 73 Prof. Dr. docent Dimitrie Mangeron Str., 700050 Iasi, Romania
3 Petru Poni Institute of Macromolecular Chemistry of Romanian Academy, Center of Advanced Research in Bionanoconjugates and Biopolymers, 41A Aleea Grigore Ghica Voda, 700487 Iasi, Romania

Abstract


-galactosidase from Aspergillus oryzae was strongly immobilized on magnetic particles functionalized with amino groups. By simple incubation without any activating agents, electrostatic interactions between amino groups and enzymes allowed obtaining a strong linkage. The immobilization efficiency was studied with the quantification of amino groups of the particles and of immobilized -galactosidase. Kinetic parameters, especially the maximal velocity Vmax and the affinity Km, were determined with two substrates, o-NPG and lactose, and compared with free enzyme values in order to evaluate the influence of our immobilization methodology on the kinetic behavior of the enzyme. Therefore, magnetic capacity of the functionalized particles allows recovering and reusing the support. Results show efficient immobilization of -galactosidase (58 g/mg of support), able to hydrolyze substrates during multiple cycles of use. Thus, magnetic particles functionalized with amino groups represent an attractive support for simple and efficient -galactosidase immobilization process.

Keywords


-galactosidase; immobilization; lactose; magnetic; particles

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