Investigation of yeast invertase immobilization onto cupric ion-chelated, porous, and biocompatible poly(hydroxyethyl methacrylate-n-vinyl imidazole) microspheres.
Cupric ion-chelated poly(hydroxyethyl methacrylate-n-vinyl imidazole) (poly(HEMA-VIM)) microspheres prepared by suspension polymerization were investigated as a specific adsorbent for immobilization of yeast invertase in a batch system. They were characterized by scanning electron microscopy, surface area, and pore size measurements. They have spherical shape and porous structure. The specific surface area of the p(HEMA-VIM) spheres was found to be 81.2 m²/g with a size range of 70-120 μm in diameter, and the swelling ratio was 86.9%. Then, Cu(II) ion chelated on the microspheres (546 μmol Cu(II)/g), and they were used in the invertase adsorption. Maximum invertase adsorption was 51.2 mg/g at pH 4.5. Cu(II) chelation increases the tendency from Freundlich-type to Langmuir-type adsorption model. The optimum activity for both free and adsorbed invertase was observed at pH 4.5. The optimum temperature for the poly(HEMA-VIM)/Cu(II)-invertase system was found to be at 55 °C, 10 °C higher than that of the free enzyme at 45 °C. V(max) values were determined as 342 and 304 U/mg enzyme, for free and adsorbed invertase, respectively. K(m) values were found to be same for free and adsorbed invertase (20 mM). Thermal and pH stability and reusability of invertase increased with immobilization.
Sari
Müfrettin Murat
MM
Biochemistry Division, Department of Chemistry, Hacettepe University, 06532 Ankara, Turkey. mufrettin@yahoo.com
D000327:Adsorption; D002614:Chelating Agents / Q000737:chemistry; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry*; D020100:Hydrogels; D007093:Imidazoles / Q000737:chemistry; D007700:Kinetics; D008863:Microspheres; D016062:Porosity; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000201:enzymology*; D013696:Temperature; D043324:beta-Fructofuranosidase / Q000737:chemistry*
D002614:Chelating Agents; D004800:Enzymes, Immobilized; D020100:Hydrogels; D007093:Imidazoles; D043324:beta-Fructofuranosidase