Glucose oxidase immobilization on a novel cellulose acetate-polymethylmethacrylate membrane.

Glucose oxidase (GOD) was immobilized on cellulose acetate-polymethylmethacrylate (CA-PMMA) membrane. The immobilized GOD showed better performance as compared to the free enzyme in terms of thermal stability retaining 46% of the original activity at 70 degrees C where the original activity corresponded to that obtained at 20 degrees C. FT-IR and SEM were employed to study the membrane morphology and structure after treatment at 70 degrees C. The pH profile of the immobilized and the free enzyme was found to be similar. A 2.4-fold increase in Km value was observed after immobilization whereas Vmax value was lower for the immobilized GOD. Immobilized glucose oxidase showed improved operational stability by maintaining 33% of the initial activity after 35 cycles of repeated use and was found to retain 94% of activity after 1 month storage period. Improved resistance against urea denaturation was achieved and the immobilized glucose oxidase retained 50% of the activity without urea in the presence of 5M urea whereas free enzyme retained only 8% activity.

[1]  Jie Wang,et al.  Characterization of cellulose acetate micropore membrane immobilized acylase I , 2004, Journal of Zhejiang University. Science.

[2]  M. Kálmán,et al.  Preparation, characterization, and potential application of an immobilized glucose oxidase , 1987, Applied biochemistry and biotechnology.

[3]  K. Siddiqui,et al.  Partial and complete alteration of surface charges of carboxymethylcellulase by chemical modification: thermostabilization in water-miscible organic solvent , 1999 .

[4]  H. Ayhan,et al.  Modified PMMA monosize microbeads for glucose oxidase immobilization , 1997 .

[5]  Y. Ikada,et al.  Lipoprotein lipase immobilization onto polyacrolein microspheres , 1990, Biotechnology and bioengineering.

[6]  J. Pazur [18] Glucose oxidase from Aspergillus niger , 1966 .

[7]  Mansoor Amiji,et al.  Enzyme immobilization in novel alginate-chitosan core-shell microcapsules. , 2004, Biomaterials.

[8]  S. Colowick,et al.  Methods in Enzymology , Vol , 1966 .

[9]  G. S. Wilson,et al.  Biosensors : fundamentals and applications , 1987 .

[10]  T. Godjevargova Behavior of glucose oxidase immobilized on ultrafiltration membranes obtained by copolymerizing acrylonitrile and N-vinylimidazol , 2000 .

[11]  Y. Yagcı,et al.  Immobilization of glucose oxidase in polypyrrole/polytetrahydrofuran graft copolymers. , 2002, International journal of biological macromolecules.

[12]  R. Tan,et al.  Immobilization of glucose oxidase on chitosan–SiO2 gel , 2004 .

[13]  Rajesh Kumar,et al.  Covalent immobilization of glucose oxidase to poly(O‐amino benzoic acid) for application to glucose biosensor , 2000 .

[14]  M. Karim,et al.  Preparation and properties of immobilized pummelo limonoid glucosyltransferase , 2002 .

[15]  J. Bailey,et al.  Immobilization of glucoamylase and glucose oxidase in activated carbon: Effects of particle size and immobilization conditions on enzyme activity and effectiveness , 1983, Biotechnology and bioengineering.

[16]  N. Karanth,et al.  Stabilization of immobilized glucose oxidase against thermal inactivation by silanization for biosensor applications. , 2004, Biosensors & bioelectronics.

[17]  M. Weibel,et al.  Pore diffusion model for a two‐substrate enzymatic reaction: Application to galactose oxidase immobilized on porous glass particles , 1976, Biotechnology and bioengineering.

[18]  V. Hasırcı,et al.  Immobilization of glucose oxidase: a comparison of entrapment and covalent bonding. , 2007, Journal of chemical technology and biotechnology.

[19]  J. Qian,et al.  Immobilization of glucose oxidase in the regenerated silk fibroin membrane: Characterization of the membrane structure and its application to an amperometric glucose sensor employing ferrocene as electron shuttle , 1995 .

[20]  Guo-Li Shen,et al.  Amperometric glucose biosensor based on a surface treated nanoporous ZrO2/Chitosan composite film as immobilization matrix , 2004 .

[21]  J. H. Lee,et al.  Immobilization of proteins on poly(methyl methacrylate) films. , 1993, Biomaterials.

[22]  T. Godjevargova,et al.  Preparation of an ultrafiltration membrane from the copolymer of acrylonitrile–glycidylmethacrylate utilized for immobilization of glucose oxidase , 1999 .

[23]  F. Kargı,et al.  Bioprocess Engineering: Basic Concepts , 1991 .

[24]  V. Bhakuni,et al.  Monovalent cation-induced conformational change in glucose oxidase leading to stabilization of the enzyme. , 2001, Biochemistry.

[25]  M. N. Gupta,et al.  A smart bioconjugate of chymotrypsin. , 2003, Biomacromolecules.

[26]  J. Lott,et al.  Evaluation of Trinder's glucose oxidase method for measuring glucose in serum and urine. , 1975, Clinical chemistry.

[27]  K. Neoh,et al.  Covalent immobilization of glucose oxidase on microporous membranes prepared from poly(vinylidene fluoride) with grafted poly(acrylic acid) side chains , 2002 .

[28]  S. Rajeswari,et al.  Development and Spectral Characterization of Poly(Methyl Methacrylate) /Hydroxyapatite Composite for Biomedical Applications , 2004 .

[29]  N. G. Karanth,et al.  Stability Studies on Immobilized Glucose Oxidase Usingan Amperometric Biosensor – Effect of Protein Based Stabilizing Agents , 2001 .

[30]  Gabriel Zoldák,et al.  Irreversible Thermal Denaturation of Glucose Oxidase from Aspergillus niger Is the Transition to the Denatured State with Residual Structure* , 2004, Journal of Biological Chemistry.

[31]  R. Landers,et al.  The ion exchange properties and equilibrium constants of Li+, Na+ and K+ on zirconium phosphate highly dispersed on a cellulose acetate fibers surface , 2004 .

[32]  S. Suye,et al.  Immobilization of glucose oxidase on poly‐(L‐lysine)‐modified polycarbonate membrane , 1998 .

[33]  Y. Yagcı,et al.  Immobilization of invertase and glucose oxidase in poly 2-methylbutyl-2-(3-thienyl) acetate/polypyrrole matrices , 2003 .

[34]  T. Scheper,et al.  Application of reversible immobilization techniques for biosensors , 1998 .