title: Immobilization of thermostable β-galactosidase on epoxy support and its use for lactose hydrolysis and galactooligosaccharides biosynthesis.
abstract: Thermoresistant, recombinant β-galactosidase from Thermotoga maritima was purified and immobilized on the surface of epoxy-coated magnetic beads. The enzyme, which has hexameric quaternary structure as shown by gel filtration chromatography, attaches to the resin through multiple covalent linkages that involve different subunits. The bound enzyme shows higher stability than the free form. The immobilized enzyme showed to be efficient for the hydrolysis of lactose and the biosynthesis of galactooligosaccharides (GOS). The chemical structure of synthesized GOS has been determined by NMR revealing that the main product was β-3′-galactosyl lactose. Although β-galactosidases from different sources have been used for the same purposes, the distinct advantage of the methodology described in this communication is that the enzyme can be easily produced, purified and immobilized in large quantities.
authors.forename: Julia
authors.initials: J
authors.lastname: Marín-Navarro
authors.forename: David
authors.initials: D
authors.lastname: Talens-Perales
authors.forename: Anneloes
authors.initials: A
authors.lastname: Oude-Vrielink
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Cañada
authors.forename: Julio
authors.initials: J
authors.lastname: Polaina
mesh_terms: D002850:Chromatography, Gel; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006868:Hydrolysis; D007785:Lactose / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D009844:Oligosaccharides / Q000096:biosynthesis*; D055503:Protein Multimerization; D021122:Protein Subunits / Q000032:analysis; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020124:Thermotoga maritima / Q000201:enzymology / Q000235:genetics; D001616:beta-Galactosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D009844:Oligosaccharides; D021122:Protein Subunits; D011994:Recombinant Proteins; D001616:beta-Galactosidase; D007785:Lactose