Immobilized metal ion affinity nanospheres for a-amylase immobilization

Authors : Tülden KALBURCU, Münire Nalan TÜZMEN

Pages : 28-40

Doi:10.3906/kim-1301-87

View : 15 | Download : 4

Publication Date : 0000-00-00

Article Type : Research Paper

Abstract :Immobilized metal chelate affinity chromatography insert ignore into journalissuearticles values(IMAC); support was practiced for a-amylase immobilization. Polyinsert ignore into journalissuearticles values(hydroxyethylmethacrylate-methacryloylamidotryptophan);-Ni2+ [pinsert ignore into journalissuearticles values(HEMA-MAT);-Ni2+] nanospheres, average diameter 100 nm, were produced by surfactant free emulsion polymerization. Characterizations of pinsert ignore into journalissuearticles values(HEMA-MAT);-Ni2+ nanospheres were carried out by Fourier transform infrared insert ignore into journalissuearticles values(FTIR); spectroscopy and scanning electron microscope insert ignore into journalissuearticles values(SEM);. In addition, average particle size, size distribution, and surface charge were specified. The amount of N-methacryloylamidotryptophan insert ignore into journalissuearticles values(MAT); incorporated to polymer was determined as 1.95 mmol/g polymers by using nitrogen stoichiometry. The specific surface areas of polyinsert ignore into journalissuearticles values(hydroxyethylmethacrylate); [pinsert ignore into journalissuearticles values(HEMA);] and pinsert ignore into journalissuearticles values(HEMA-MAT); nanospheres were calculated as 1856 m2/g and 1914 m2/g, respectively. Protein adsorption increased with increasing initial protein concentration and maximum a-amylase adsorption on pinsert ignore into journalissuearticles values(HEMA-MAT);-Ni2+ nanospheres was observed at pH 4.0. Both free and immobilized a-amylase showed pH optimum at pH 7.0. It was determined that the immobilized a-amylase had better thermostability than the free one. Immobilization of the enzyme did not significantly change the kinetic parameters. The storage stability of a-amylase increased upon immobilization. It was also observed that pinsert ignore into journalissuearticles values(HEMA-MAT);-Ni2+ nanospheres can be repeatedly used for a-amylase immobilization.
Keywords : a Amylase, nanospheres, IMAC, enzyme immobilization, adsorption