Influence of green solvents on enzymatic activity of free and inmobilized b-galactosidases from Bacillus circulans

The oligosaccharide present on the cell surface are involved in many biological processes such as cell-cell recognition and communication, growth regulation and antibody interaction [1]. Nowadays there is an increasing need to obtain these compounds for biological studies but their chemical synthesis is still a time-consuming approach, often expensive and using toxic catalysts. The use of glycosidase provides an alternative approach for the preparation of oligosaccharides with biological activity [2]. At the same time, the use of enzymes in Ionic liquids (ILs) has proven to have many advantages, such as, high conversion rates, high enantioselectivity, and better enzyme stability as well as better recoverability and recyclability [3]. Biolacta N\uba 5 it\u2019s a crude preparation of b-galactosidases from Bacillus circulans that exhibits high yields in the synthesis of Gal-b-(1\u21924)GlcNac and a very low rates of Gal-b-(1\u21926)GlcNac, using pNP-b-Gal as donor and GlcNac as acceptor [4]. b\u2013gal-3 is a pure b\u2013galactosidase from Bacillus circulans (strain ATC 31382) which exhibits high yields in the synthesis of Gal-b-(1\u21923)GlcNac and Gal-b-(1\u21923)GalNAc using pNP-b-Gal as donor and GlcNac or GalNAc as acceptor. In this comunication we have studied the influence of different green solvents (ILs and solvents from biomass) as co-solvents in the enzymatic synthesis of disaccharides using these two b-galactosidases; Biolacta and b-gal-3 cloned in our laboratory. Some of these green solvents change the classical regioselectivity of these enzymes. We also investigated the covalent immobilization of these b\u2013galactosidases on new porous polymers functionalized with epoxy groups. After this study, the best immobilized derivative was used for disaccharides synthesis using green solvents