title: Kinetics study of invertase covalently linked to a new functional nanogel.
abstract: Nanogels are promising materials as supports for enzyme immobilization. A new hydrogel comprising of methacrylic acid (MAAc) and N-vinyl pyrrolidone (N-VP) and ethyleneglycol dimethacrylate (EGDMA) was synthesized and converted to nanogel by an emulsification method. Nanogel was further functionalized by Curtius azide reaction for use as support for the covalent immobilization of invertase (Saccharomyces cerevisiae). As-prepared or invertase-immobilized nanogel was characterized by FTIR, XRD, TEM and nitrogen analysis. The characterization of both free and the immobilized-invertase were performed using a spectrophotometric method at 540 nm. The values of V(max), maximum reaction rate, (0.123 unit/mg), k(m), Michaelis constant (7.429 mol/L) and E(a), energy of activation (3.511 kj/mol) for the immobilized-invertase are comparable with those of the free invertase at optimum conditions (time 70 min, pH 6.0 and temperature 45°C). The covalent immobilization enhanced the pH and thermal stability of invertase. The immobilized biocatalyst was efficiently reused up to eight cycles.
authors.affiliation: Himachal Pradesh University, Department of Chemistry, Shimla, India.
authors.forename: Lok
authors.initials: L
authors.lastname: Raj
authors.forename: Ghanshyam S
authors.initials: GS
authors.lastname: Chauhan
authors.forename: Wamik
authors.initials: W
authors.lastname: Azmi
authors.forename: J-H
authors.initials: JH
authors.lastname: Ahn
authors.forename: James
authors.initials: J
authors.lastname: Manuel
mesh_terms: D004789:Enzyme Activation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry; D005782:Gels / Q000737:chemistry*; D007700:Kinetics; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D011108:Polymers / Q000737:chemistry*; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D043324:beta-Fructofuranosidase / Q000737:chemistry*
chemical_list: D004800:Enzymes, Immobilized; D005782:Gels; D011108:Polymers; D043324:beta-Fructofuranosidase