Single-step High Fructose Syrup Production from Chicory Inulin by Immobilized Inulinase on Functionalized nanomagnetite Via Protein Nanoparticles

Document Type : Research Article

Authors

1 MS in Agricultural Engineering-Food Science and Technology, Department of Agriculture Research, Iranian Research Organization for Science and Technology (IROST)

2 Assistant Prof. in Food Science and Technology, Department of Chemical Technology , Iranian Research Organization for Science and Technology (IROST)

3 Ph.D. in Analytical Chemistry, Young Researchers and Elite Club, North Tehran Branch, Islamic Azad University, Tehran, Iran

Abstract

 In this study, Inulinase enzyme was immobilized on the magnetite nanoparticles functionalized with protein nanoparticles through covalent attachment to improve the stability of the enzyme. The Inulinase was immobilized in order to increase enzyme stability under extreme conditions of industrial processes, reusability, reducing the cost of the final products and the ability to use of continuous bioreactors with immobilized enzyme. In addition, the immobilized enzyme on magnetite nanoparticles can separate from the reaction medium by the magnetic field. Therefore, at the first step, the magnetite nanoparticles were prepared by co-precipitation method. Then, in the second step, a mixture of soy protein isolate and bovine serum albumin nanoparticles were synthesized by desolvation method and then the magnetite nanoparticle’s surface were coated for modification and stabilization. In The third step, the enzyme was immobilized on the surface of the magnetite carrier. In this study, scanning electron microscope (SEM) and dynamic light scattering (DLS) were used for analyze the particle size, shape, morphology and functional characteristics of magnetite and protein nanoparticles and enzyme immobilization in all steps of preparation. After enzyme immobilization the properties were compared with the free enzyme. The results implied that the immobilized enzyme retained more than 50% of its initial activity at 650C while the free enzyme retained only 17% of its activity. The half-life of immobilized enzyme at 650C, increased up to 6 times in comparison with the free enzyme. The immobilized inulinase retained more than 60% of its activity after 12 cycles. The activity of the enzyme, kinetic parameters (Vmax, Km) were assayed and the results revealed better operational stability compared to free enzyme.

Keywords

Main Subjects

References

[1] Nakamura, T., Ogata Y., Akichika S., Nakamura A., Ohta K. (1995). Continuous production of fructose syrups from inulin by immobilized inulinase from Aspergillus niger mutant 817. J. Ferment. Bioeng., 80, 164-169.
[2] Datta, S., Christena, L.R., Rani, Y., Rajaram, S. (2012). Enzyme immobilization: An verview on techniques and support materials. Biotechnol., 3, 1-9.
[3] Liang,  X., Shi, H., Jia, X., Yang, Y., Liu, X. (2011). Dispersibility, shape and magnetic properties of nano-Fe3O4 particles. Mater. Sci. Appl., 2, 1644-1653.
[4] Gupta, A., Gupta, M. (2005). Synthesis and surface engineering of iron oxide nanoparticles for biomedical applications. Biomaterials, 26, 3995-4021.
[5] Reddy, L., Arias, J., Nicolas, J., Couvreur, P. (2012). Magnetic nanoparticles: Design and Characterization, Toxicity and Biocompatibility, Pharmaceutical and Biomedical Applications. Chem. Rev., 112, 5818-5878.
[6] Lohcharoenkal, W., Wang, L., Chen, Y.C., Rojanasakul, Y. (2014). Protein nanoparticles as drug delivery carriers for cancer therapy. J. Biomed. Biotechnol., 2014, 1-15.
[7] Sundar, S., Kundu, J., Kundu, S.C. (2010). Biopolymeric nanoparticles. Sci. Tech. Adv. Mater., 11, 1-13.
[8] Kim, W.Y., Byun, S.M., Uhm, T.B. (1982). Hydrolysis  of  inulin  from  Jerusalem  artichoke  by  inulinase  immobilized  on aminoethylcellulose. Enzyme Microb. Technol., 4, 239-244.
[9] Ettalibi, M., Baratti, J.C. (1992). Immobilization of Aspergillus ficuum inulinases on porous glass. Biocatal. Biotransformation, 5, 175-182.
[10] Kim, D.H., Choi, Y.J., Song, S.K., Yun, J.W. (1997). Production of inulo-oligosaccharides using endo-inulinases from a Pseudomonas sp. Biotechnol. Lett., 19, 369-371.
[11] Kovaleva, T.A., Holyavka, M.G., Bogdanova, S.S. (2009). Inulinase immobilization on macroporous anion-exchange resins by different methods. Bull. Exp. Biol. Med., 148, 39-41.
[12] Elnashar, M.M., Danial, E.N., Awad, G.E. (2009). Novel carrier of grafted alginate for covalent immobilization of inulinase. ‎Ind. Eng. Chem. Res., 48, 9781-9785.
[13] Nguyen, Q.D., Rezessy, J.M., Czukor, B., Hoschke, A. (2011). Continuous production of oligofructose syrup from Jerusalem artichoke juice by immobilized endo-inulinase. Process Biochem. , 46, 298-303.
[14] Zemolin, G.P., Gazoni, M., Zabot, G.L., Golunski, S.M., Astolfi, V., Pra, V.D. (2012). Immobilization of inulinase obtained by solid-state fermentation using spray-drying technology. Biocatal. Biotransformation, 30, 409-416.
[15] Coghetto, C.C., Scherer, R.P., Silva, M.F., Golunski, S., Pergher, S.B.C., Oliveira, D.D.,  Oliveira, J.V.,  Treichel, H. (2012). Natural montmorillonite as support for the immobilization of inulinase from Kluyveromyces marxianus NRRL Y-7571. Biocatal. Agric. Biotechnol., 1, 284-289.
[16] Yewale, T., Singhal, R.S., Vaidya, A.A. (2013). Immobilization of inulinase from Aspergillus niger NCIM 945 on chitosan and its application in continuous inulin hydrolysis. Biocatal. Agric. Biotechnol., 2, 96-101.
[17] Missau, J., Scheid, A.J., Foletto, E.L., Jahn, S.L., Mazutti, M.A., Kuhn, R.C. (2014). Immobilization of commercial inulinase on alginate–chitosan beads. Sustain. Chem. Process., 2, 2-13.
[18] Torabizadeh, H.,  Tavakoli, M., Safari, M. (2014). Immobilization of thermostable Alpha-amylase from Bacillus licheniformis by cross-linked enzyme aggregates method using calcium and sodium ions as additives. J. Mol. Catal. B-Enzym., 108, 13-20.
[19] Teng, Z., Luo, Y., Wang, Q. (2012). Nanoparticles synthesized from soy protein: Preparation, characterization, and application for nutraceutical encapsulation. J. Agric. Food Chem., 60, 2712-2720.
[20] Zhanfeng, L., Linhui, Q., Shuangling, Z., Hongyan, W., Xuejun, C. (2013). Synthesis  and  characterization  of  monodisperse  magnetic  Fe3O4@BSA core–shell  nanoparticles. Colloids Surf. A Physicochem. Eng., 436, 1145-1151.
[21] Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72, 248–254.
Innovative Food Technologies
Volume 4, Issue 1
November 2016
Pages 79-91

Files

History

  • Receive Date: 05 July 2016
  • Revise Date: 10 August 2016
  • Accept Date: 06 September 2016

Share

How to cite

Statistics