Hostname: page-component-78c5997874-v9fdk Total loading time: 0 Render date: 2024-11-05T05:45:13.242Z Has data issue: false hasContentIssue false
  • English
  • Français

Immobilization of α-amylase onto K-10 montmorillonite: characterization and comparison of activity in a batch and a fixed-bed reactor

Published online by Cambridge University Press:  09 July 2018

G. Sanjay  and
S. Sugunan
G. Sanjay
Affiliation:
Department of Applied Chemistry, Cochin University of Science and Technology, Kochi - 682022, India
S. Sugunan*
Affiliation:
Department of Applied Chemistry, Cochin University of Science and Technology, Kochi - 682022, India
*
E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

α-amylase was immobilized on acid-activated montmorillonite K-10 via adsorption and covalent linkage. The immobilized enzymes were characterized by X-ray diffraction (XRD), surface area measurements, 27Al nuclear magnetic resonance (NMR) and scanning electron microscopy (SEM). Surface area measurements indicate pore blockage due to linking of the enzyme in the vicinity of the pore mouth. The XRD demonstrates intercalation of enzyme upon immobilization. The NMR studies indicate that, during adsorption, tetrahedral Al sites are involved, while covalent binding occurs exclusively on the octahedral Al sites. The SEM images depict the changed morphology of the clay surface due to immobilization. The efficiency of immobilized enzymes for starch hydrolysis was tested in a batch and a fixed-bed reactor and the performances were compared. The immobilized α-amylase showed a broad pH profile and improved stability characteristics in both reactor types when compared to the free enzyme. The effectiveness factor increased in the fixed-bed reactor, implying that diffusional restrictions to mass transfer operate in the heterogeneous reaction and the use of a fixed-bed reactor leads to a reduction in these diffusional resistances. In the continuous run, 100% initial activity was maintained for 72 h, and after 96 h, >80% activity was retained.

Keywords

α-amylasemontmorilloniteadsorptioncovalent bindingimmobilized enzymesstarch hydrolysis
Type
Research Article
Information
Clay Minerals , Volume 40 , Issue 4 , December 2005 , pp. 499 - 510
Copyright
Copyright © The Mineralogical Society of Great Britain and Ireland 2005

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Akgol, S., Kacar, Y., Denizli, A. & Arica, M.Y. (2001) Hydrolysis of sucrose by invertase immobilized onto novel magnetic polyvinylalcohol microspheres. Food Chemistry, 74, 281–288.Google Scholar
Aksoy, S., Tumturk, H. & Hasirci, N. (1998) Stability of a-amylase immobilized on poly (methyl metacrylate-acrylic acid) microspheres. Journal of Biotechnology, 60, 37–46.Google Scholar
Arica, M.Y. & Hasirci, V. (1993) Immobilization of glucose oxidase: A comparison of entrapment and covalent binding. Journal of Chemical Technology and Biotechnology, 58, 287–292.Google Scholar
Arica, M.Y., Hasirci, V. & Alaeddinoglu, N.G. (1995) Covalent immobilization of a-amylase onto α-HEMA microspheres: preparation and application to fixed bed reactor. Biomaterials, 15, 761–768.Google Scholar
Balasubramaniam, K. & Arasaratnam, V. (1989) Kinetic studies on soluble and immobilized alpha amylase and glucoamylase. Journal of the National Science Council Sri Lanka, 17, 91–97.Google Scholar
Bayramoglu, G., Yilmaz, M. & Arica, M.Y. (2004) Immobilization of a thermostable a-amylase onto reactive membranes: kinetics characterization and application to continuous starch hydrolysis. Food Chemistry, 84, 591–599.CrossRefGoogle Scholar
Chen, J.P., Chu, D.H. & Sun, Y.M. (1997) Immobilization of a-amylase to temperature responsive polymers by single or multiple point attachment. Journal of Chemical Technology and Biotechnology, 69, 421–428.Google Scholar
Gaikwad, S.M. & Deshpande, V.V. (1992) Immobilization of glucose isomerase on Indion 48R. Enzyme and Microbial Technology, 14, 855–858.CrossRefGoogle Scholar
arwood, G.A., Mortland, M.M. & Pinnavaia, T.J. (1983) Immobilization of glucose oxidase on montmorillo- nite clay: hydrophobic and ionic modes of binding. Journal of Molecular Catalysis, 22, 153–163.Google Scholar
Gougeon, R.D., Soulard, M., Reinholdt, M., Brendle, J.- M., Chezeau, R., LeDred, R., Marchal, R. & Jeandet, P. (2003) Polypeptide adsorption on a synthetic montmorillonite: A combined solid-state NMR spectroscopy, X-ray diffraction, thermal analysis and N2 adsorption study. European Journal of Inorganic Chemistry, 1366–1372.Google Scholar
Handa, T., Hirose, A., Akino, T., Watanabe, K. & Tsuchiya, H. (1983) Preparation of immobilized α- amylase covalently attached to granular polyacrylonitrile. Biotechnology and Bioengineering, 25, 2957–2967.Google Scholar
Hess, J.M. & Kelly, R.M. (1999) Influence of polymolecular events on the inactivation behaviour of Xylose isomerase from thermotoga neapolitana 5068. Biotechnology and Bioengineering, 62 509–517.3.0.CO;2-7>CrossRefGoogle Scholar
Jia, W., Segal, E., Kornemandal, D., Lamhot, Y., Narkis, M. & Siegmann, A. (2002) Polyaniline-DBSA/ organophilic clay nanocomposites: synthesis and characterization. Synthetic Metals, 128, 115–120.CrossRefGoogle Scholar
Ju, Y., Chen, W. & Lee, C. (1995) Starch slurry hydrolysis using a-amylase immobilized on a hollow fiber reactor. Enzyme Microbial Technology, 17, 685–688.Google Scholar
Kim, B.H., Jung, J.H., Hong, S.H., Kim, J.W., Choi, H.J. & Joo, J. (2001) Physical characterization of emulsion intercalated polyaniline-clay nanocomposites. Current and Applied Physics, 1, 112–115.CrossRefGoogle Scholar
Lai, C.M. & Tabatabai, M.A. (1992) Kinetic parameters of immobilized urease. Soil Biology and Biochemistry, 24, 225–228.CrossRefGoogle Scholar
Laszlo, P. (1987) Chemical reactions on clays. Science, 23, 235–239.Google Scholar
Lee, P.M., Lee, K.H. & Siaw, S.Y. (1993) Covalent immobilization of aminoacrylase to alginate for 2-phenyl alanine production. Journal of Chemical Technology and Biotechnology, 58, 65–70.Google Scholar
Lim, L.H., Macdonald, D.G. & Hill, G.A. (2003) Hydrolysis of starch particles using barley a-amylase. Biochemical Engineering Journal, 13, 53–62.Google Scholar
Lomako, O.V., Menyailova, I.I., Nakhapetyan, L.A., Nikitin, Y. & Kiselev, A.V. (1982) Immobilization of glucoamylase on porous silicas. Enzyme and Microbial Technology, 4, 89–92.Google Scholar
Lowry, O.H., Rosebrough, N.J., Faar, A.L. & Randall, R.J.J. (1951) Protein measurement with the folin phenol reagent. Journal of Biological Chemistry, 193, 265–275.Google Scholar
Mody, H.M., Mody, K.H., Mairh, O.P. & Jasra, R.V. (1999) Immobilization of alpha amylase on porous silica. Indian Journal of Chemistry, 38A, 1200–1202.Google Scholar
Naidja, A. & Huang, P.M. (1996) Deamination of aspartic acid by aspartase-Ca-montmorillonite complex. Journal of Molecular Catalysis A: Chemical, 106, 255–265.CrossRefGoogle Scholar
Naidja, A., Huang, P.M. & Bollag, J.-M. (1997) Activity of tyrosinase immobilized on hydroxyaluminum- montmorillonite complexes. Journal of Molecular Catalysis A: Chemical, 115, 305–316.CrossRefGoogle Scholar
Noda, T. & Suda, S.F. (2001) Sweet potato a-amylase immobilized on chitosan beads and its application in a semi continuous production of maltose. Carbohydrate Polymers, 44, 189–195 Google Scholar
Pinnavaia, T.J. (1983) Intercalated clay catalysts. Science, 220, 365–371.Google Scholar
Sarkar, J.M., Leonowicz, A. & Bollag, J.-M. (1989) Immobilization of enzymes on clays and soils. Soil Biology and Biochemistry, 21, 223–230.CrossRefGoogle Scholar
Silverman, R.B. (2000) The Organic Chemistry of Enzyme-Catalyzed Reactions. Academic Press, USA, 569 pp.Google Scholar
Siso, M.I.G., Graber, M., Condoret, J.-S. & Combes, D. (1990) Effect of diffusional resistances on the action pattern of immobilized alpha amylase. Journal of Chemical Technology and Biotechnology, 48, 185–200.Google ScholarPubMed
Synowiecki, J., Sikorski, Z.E., Caczk, M. & Piotrzkowska, H. (1982) Immobilization of enzymes on Krill Chitin activated by formaldehyde. Biotechnology and Bioengineering, 24, 1871–1876.CrossRefGoogle ScholarPubMed
Tanyolac, D., Yuruksoy, B.I. & Ozdural, A.R. (1998) Immobilization of a thermostable a-amylase, Thermamyl, onto nitrocellulose membrane by Cibracon Blue F3GA dye binding. Biochemical Engineering Journal, 2, 179–186.CrossRefGoogle Scholar
Tumturk, H., Aksoy, S. & Hasirci, N. (2000) Covalent immobilization of a-amylase onto poly (2-hydroxy- methyl methacrylate) and poly (styrene-2-hydroxy- methyl methacrylate) microspheres and the effect of Ca2+ ions on the enzyme activity. Food Chemistry, 68, 259–266.Google Scholar
Ulbrich, R., Schellenberger, A. & Damerau, W. (1986) Studies on thermal inactivation of immobilized enzymes. Biotechnology and Bioengineering, 28, 511–522.Google Scholar