Abstract
Carboxymethyl cellulose-silver nanoparticle (AgNp)-silica hybrids have been synthesized in a modified Stöber process. The hybrid synthesis was optimized to obtain an efficient immobilization matrix for diastase alpha amylase, a multimeric enzyme of high technological significance. The synthesized hybrids were characterized using FTIR, XRD, SEM, TGA and BET studies. The enzyme immobilization was done by adsorption and using the immobilized enzyme, the hydrolysis of soluble starch has been optimized in comparison to free enzyme. The optimum usable pH for the immobilized enzyme ranged from pH 4 to 5, while pH 5 was optimum pH for the free enzyme activity. The kinetic parameters for the immobilized, (K M = 3.4610 mg ml−1; V max = 6.3540 mg ml−1 min−1) and free enzyme (K M = 4.1664 mg ml−1; V max = 4.291 mg ml−1 min−1) hydrolysis indicated that the immobilization at the nanohybrid has significantly improved the catalytic property of the enzyme. In the immobilized state, the enzyme remained usable for many repeated cycles like our previous material, gum acacia-gelatin-AgNp-silica. Storage experiments indicated that the immobilization has increased the stability of the enzyme and also that AgNps play a role in stabilizing the immobilized enzyme.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bakunina Yu, Nedashkovskaya OI, Zvyagintseva TN, Shchipunov YuA (2006) Immobilization of α-galactosidase inside hybrid silica nanocomposites containing polysaccharides. Russ J Appl Chem 79:827
Benmouhoub N, Simmonet N, Agoudjila N, Coradin T (2008) Aqueous sol-gel routes to bio-composite capsules and gels. Green Chem 10:957
Copello GJ, Mebert AM, Raineri M, Pesenti MP, Diaz LE (2011) Removal of dyes from water using chitosan hydrogel/SiO2 and chitin hydrogel/SiO2 hybrid materials obtained by sol-gel method. J Hazard Mater 186:932
Djabali D, Belhaneche N, Nadjemi B, Dulong V, Picton L (2009) Relationship between potato starch isolation methods and kinetic parameters of hydrolysis by free and immobilised α-amylase on alginate (from Laminaria digitata algae). J Food Compos Anal 22:563
European Pharmacopeia, 5.0 (2005), Council of Europe (COE)—European Directorate for the Quality of Medicines (EDQM), Chapter 4, Reagents, section 4.1.3., Buffer solutions, p 430
Hebeish AA, El-Rafie MH, Abdel-Mohdy FA, Abdel-Halim ES, Emam HE (2010) Carboxymethyl cellulose for green synthesis and stabilization of silver nanoparticls. Carbohydr Polym 82:933
Hu R, Lin L, Liu T, Ouyang P, He B, Liu S (2008) Reducing sugar content in hemicellulose hydrolysate by DNS method: a revisit. J Biobased Mater Bioener 2:156
Lai S-M, Yang Ar J-M, Chen W-C, Hsiao J-F (2006) The properties and preparation of chitosan/silica hybrids using sol-gel process. Polym-Plast Technol Eng 45:997
Lee D, Cohen RE, Rubner MF (2005) Antibacterial properties of Ag nanoparticle loaded multilayers and formation of magnetically directed antibacterial microparticles. Langmuir 21:9651
Manelius R, Bertoft E (1996) The effect of Ca2+-ions on the α-amylolysis of granular starches from oats and waxy-maize. J Cereal Sci 24:139
Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426
Mohan YM, Raju KM, Sambasivudu K, Singh S, Sreedhar B (2007) Preparation of acacia-stabilized silver nanoparticles: a green approach. J Appl Polym Sci 106:3375
Rao YN, Banerjee D, Datta A, Das SK, Guin R, Saha A (2010) Gamma irradiation route to synthesis of highly re-dispersible natural polymer capped silver nanoparticles. Radiat Phys Chem 79:1240
Rojas IA, Slunt JB, Grainger DW (2000) Polyurethane coatings release bioactive antibodies to reduce bacterial adhesion. J Control Rel 63:75
Shchipunov YA, Karpenko TY (2004) Hybrid polysaccharide-silica nanocomposites prepared by the sol-gel technique. Langmuir 20:3882
Shchipunov YA, Karpenko TY, Bakunina IY, Burtseva YV, Zvyagintseva TN (2004) A new precursor for the immobilization of enzymes inside sol-gel-derived hybrid silica nanocomposites containing polysaccharides. J Biochem Biophys Meth 58:25
Shchipunov YA, Karpenko TY, Krekoten AV (2005) Hybrid organic-inorganic nanocomposites fabricated with a novel biocompatible precursor using sol-gel processing. Compos Interface 11:587
Singh V, Ahmed S (2012) Silver nanoparticle (AgNPs) doped gum acacia-gelatin-silica nanohybrid for diastase immobilization. Int J Biol Macromol 50:353
Singh V, Kumar P (2011a) Carboxymethyl tamarind gum-silica nanohybrids for effective immobilization of amylase. J Mol Catal B Enzym 70:67
Singh V, Kumar P (2011b) Design of nanostructured tamarind seed kernel polysaccharide-silica hybrids for mercury (II) removal. Sep Purif Technol 46:825
Singh V, Singh SK (2011) Synthesis and characterization of gum acacia inspired silica hybrid xerogels for mercury(II) adsorption. Inter J Biol macromol 48:445
Singh V, Singh SK, Pandey S, Sanghi R (2010) Adsorption behavior of potato starch-silica nanobiocomposite. Adv Mater Lett 1:40
Siso MIG, Graber M, Condoret J-S, Combes D (1990) Effect of diffusional resistances on the action pattern of immobilized alpha-amylase. J Chem Technol Biotechnol 48:185
Tischer W, Wedekind F (1999) Immobilized enzymes: methods and applications. Biocatalysis—From Discovery to Application. In: Fessner W-D (ed) Topics in Current Chemistry, Springer, Berlin Heidelberg, Vol. 200, pp 95–126
Travan A, Pelillo C, Donati I, Marsich E, Benincasa M, Scarpa T, Semeraro S, Turco G, Gennaro R, Paoletti S (2009) Non-cytotoxic silver nanoparticle-polysaccharide nanocomposites with antimicrobial activity. Biomacromolecules 10:1429
Wang G-H, Zhang L-M (2006) Using novel polysaccharide-silica hybrid material to construct an amperometric biosensor for hydrogen peroxide. J Phys Chem B 110:24864
Acknowledgments
The authors are grateful to the University Grants Commission, New Delhi India for the financial support to carry out this research work. IR, SEM and XRD facilities are acknowledged to Indian Institute of Mines, Dhanbad, Indian Institute of Technology Karagpur and National Centre of Experimental Mineralogy and Petrology, University of Allahabad respectively. Authors thank Dr. Anjana Pandey, Centre for Biotechnology, University of Allahabad, India for gifting dialysis bags.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Singh, V., Ahmad, S. Synthesis and characterization of carboxymethyl cellulose-silver nanoparticle (AgNp)-silica hybrid for amylase immobilization. Cellulose 19, 1759–1769 (2012). https://doi.org/10.1007/s10570-012-9749-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10570-012-9749-6