Inulinase Immobilization on Functionalized Magnetic Nanoparticles Prepared with Soy Protein Isolate Conjugated Bovine Serum Albumin for High Fructose Syrup Production
Inulinase from Aspergillus niger was covalently immobilized on magnetic nanoparticles (MNPs/Fe3O4) covered with soy protein isolate (SPI/Fe3O4) functionalized by bovine serum albumin (BSA) nanoparticles. MNPs are promising enzyme carriers because they separate easily under external magnetic fields and have enhanced immobilized enzyme reusability. As MNPs aggregate simply, surface coating strategy was employed. SPI functionalized by BSA was a suitable candidate for nanomagnetite coating due to its superior biocompatibility and hydrophilicity. Fe3O4@SPI-BSA nanoparticles were synthesized as a novel carrier with narrow particle size distribution. Step by step fabrication monitoring of Fe3O4@SPI-BSA nanoparticles was performed using field emission scanning electron microscopy and dynamic light scattering. The results illustrated that nanomagnetite with the spherical morphology was well monodispersed with the diameter of about 35 nm. The average size of the SPI-BSA nanoparticles was 80 to 90 nm, and their zeta potential was around −34 mV. Finally, the mean diameter of fabricated Fe3O4@SPI-BSA NPs was less than 120 nm. Inulinase enzyme from Aspergillus niger was covalently immobilized through gluteraldehyde on Fe3O4@SPI-BSA nanoparticles successfully. Fourier transform infrared spectra and field emission scanning electron microscopy images provided sufficient proof for the enzyme immobilization on the nanoparticles with 80% enzyme loading.
 R. Couvreur, R. Gref, K. Andrieux, K, and C. Malvy, C, “Nanotechnologies for drug delivery: Application to cancer and autoimmune diseases,” Prog. Solid State Chem., vol. 34, pp. 231-235, Jul. 2006.
 S. Kumaresh, T. M Aminabhavi, A. R Kulkarni, and W. E Rudzinski, “Biodegradable polymeric nanoparticles as drug delivery devices,” J. Control. Release., vol. 70, pp. 1-20, Jan. 2001.
 T. Xie, A. M Wang, L. F Huang, H. F Li, Z. M Chen, and Q. Y Wang, “Recent advance in the support and technology used in enzyme immobilization,” Afr. J. Biotechnol., vol. 8, pp. 4724–4733, Oct. 2009.
 H. Jia, G. Zhu, and P. Wang, “Catalytic behaviors of enzymes attached to nanoparticles: the effect of particle mobility,” Biotechnol. Bioeng., vol. 84, pp. 406-414, Nov. 2003.
 S. Datta, L. R. Christena, Y. Rani, and S. Rajaram, “Enzyme immobilization: An review on techniques and support materials,” Biotechnol., vol. 3, pp. 1-9, Feb. 2012.
 R. A. Sheldon, “Enzyme immobilization: The quest for optimum performance,” Adv. Synth. Catal., vol. 349, pp. 1289-1307, Jun. 2007.
 H. Torabizadeh, M. Tavakoli, and M. Safari, “Immobilization of thermostable α-amylase from Bacillus licheniformis by cross-linked enzyme aggregates method using calcium and sodium ions as additives,” J. Mol. Catal. B: Enzym., vol.108, pp. 13-20, Jun. 2014.
 A. Richetti, C. B. Munaretto, L. A. Lerin, L. Batistella, J. V. Oliveira, R. M. Dallago, V. Astolfi, M. D. Luccio, M.A. Mazutti, D. D. Oliveira, and H. Treichel, “Immobilization of inulinase from Kluyveromyces marxianus NRRL Y-7571 using modified sodium alginate beads,” Bioprocess and Biosys. Eng., vol. 35, pp. 383-388, Aug. 2012.
 D. T. Mitchell, S. B. Lee, L. Trofin, N. Li, T. K. Nevanen, H. Soderlund, C. R. and martin, “Smart nanotubes for bioseparations and biocatalysis,” J. Am. Chem. Soc., vol. 124, pp. 11864–11865, Oct. 2002.
 L. H. Reddy, J. Arias, J. Nicolas, and P. Couvreur, “Magnetic nanoparticles: Design and Characterization, Toxicity and Biocompatibility. Pharmaceutical and Biomedical Applications,” Chem. Rev., vol. 112, pp. 5818-5878, Oct. 2012.
 G. Couto, J. Klein, W. Schreiner, D. Mosca, A. Oliveira, and A. Zarbin, “Nickel nanoparticles obtained by a modified polyol process: Synthesis, Characterization, and Magnetic Properties,” J. Colloid Interface Sci., vol. 311, pp. 461-468, Jul. 2007.
 C. Sun, J. Lee, and M. Zhang, “Magnetic nanoparticles in MR imaging and drug delivery. Adv. Drug Deliv. Rev., vol. 60, pp. 1252-1265, Apr. 2008.
 X. Liang, H. Shi, X. Jia, Y. Yang, and X. Liu, “Dispersibility, shape and magnetic properties of nano-Fe3O4 particles,” Mater. Sci. Appl., vol. 2, pp. 1644-1653, Mar. 2011.
 A. Gupta, and M. Gupta, “Synthesis and surface engineering of iron oxide nanoparticles for biomedical applications,” Biomaterials., vol. 26, pp. 3995-4021, Jun. 2005.
 M. Mikhaylova, D. K. Kim, C. Catherine, and S. G. Adam, “BSA immobilization on amine-functionalized superparamagnetic iron oxide nanoparticles,” Chem. Mater., vol. 16, pp. 2344-2354, Sep. 2004.
 L. Zhanfeng, Q. Linhui, Z. Shuangling, W. Hongyan, and C. Xuejun, “Synthesis and characterization of monodisperse magnetic Fe3O4@BSA core–shell nanoparticles,” Colloids Surf., vol. 436, pp. 1145-1151, Sep. 2013.
 I. Nedkov, T. Merodiiska, L. Slavov, R. E. Vandenberhe, Y. Kusano, and J. Takada J, “Surface oxidation, size and shape of nano-sized magnetite obtained by co-precipitation,” J. Magn. Magn. Mater., vol. 300, pp. 358-367, Nov. 2006.
 H. Itoh, and T. Sugimoto, “Systematic Control of Size, Shape, Structure, and Magnetic Properties of Uniform Magnetite and Maghemite Particles,” J. Colloid. Interface. Sci., vol. 12, pp. 283–295, Sep. 2003.
 M. C. Mascolo, Y. Pei, and T. A. Ring, “Room temperature co-precipitation synthesis of magnetite nanoparticles in a large pH window with different bases,” Materials., vol. 6, pp. 5549-5567, Nov. 2013.
 N. D. Kandpal, S. Sah, R. Loshali, R. Joshi, and J. Prasad, “Copercipitation method of characterization of iron oxide nanoparticles,” J. Sci. Ind. Res., vol. 73, pp. 87-90, Feb. 2014.
 W. Lohcharoenkal, L. Wang, Y. C. Chen, and Y. Rojanasakul, “Protein nanoparticles as drug delivery carriers for cancer therapy,” J. Biomed. Biotechnol., vol. 2014, pp. 1-15, Jan. 2014.
 C. Weber, C. Coester, J. Kreuter and K. Langer, “Desolvation process and surface characterization of protein nanoparticles,” Int. J. Pharm., vol. 194, pp. 91-102, Jan. 2000.
 Z. Teng, Y. Luo, and Q. Wang, “Nanoparticles synthesized from soy protein: Preparation, characterization, and application for nutraceutical encapsulation,” J. Agric. Food. Chem., vol. 60, pp. 2712-2720, Mar. 2012.
 S. A Krishna, P. Amareshwar, and P. Chakravarty, “Different techniques used for the preparation of nanoparticles using natural polymers and their application,” J. Pharm. Pharm. Sci., vol. 3, pp. 45-50, Oct. 2011.
 J. Y. Jun, H. H. Nguyen, S. Y. R. Paik, H. S. Chun, B. C. Kang, and S. Ko, “Preparation of size-controlled bovine serum albumin (BSA) nanoparticles by a modified desolvation method,” Food. Chem., vol. 4, pp. 1892-1898, Dec. 2011.
 F. Galisteo-González and J. A. Molina-Bolívar, “Systematic study on the preparation of BSA nanoparticles,” Colloids Surf., B., vol. 123, pp. 286-292 Nov. 2014.
 R. Mehravar, M. Jahanshahi, and N. Saghtoleslami, “Fabrication and evaluation of human serum albumin (HAS) nanoparticles for drug delivery application,” Int. J. Nanosci., vol. 8, pp. 319–322, Nov. 2009.
 M. Jahanshahi, G. D. Najafpour, and M. Rahimnejad, “Applying the Taguchi method for optimized fabrication of bovine serum albumin (BSA) nanoparticles as drug delivery vehicles,” Afr. J. Biotechnol., vol. 7, pp. 362-367, Feb. 2008.
 D. A. Skoog, F. J. Holler, and S. R. Crontch, Principles of instrumental analysis, Thomson Brooks/Cole, Canada, 2007, pp. 237-250.
 P. Singh, P. K. and Gill, “Production of inulinases: Recent advances,” Biotechnol., vol. 44, pp. 151-162, Feb. 2006.
 T. M. Mohamed, S. M. El-Souod, E. M. Ali, M. O. El-Badry, and M.M. El-Keiy, “Immobilization and characterization of inulinase from Ul ocladium atrumon nonwoven fabrics,” J. Biosci., vol. 39, pp. 785–793, Nov. 2014.
 M. M. Bradford, “A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding,” Anal. Biochem., vol. 72, pp. 248–254, May. 1976.
 H. Torabizadeh, M. Habibi-Rezaei, M. Safari, A. A. Moosavi-Movahedi, and H. Razavia, “Semi-rational chemical modification of endoinulinase by pyridoxal 5′-phosphate and ascorbic acid,” J. Mol. Catal. B: Enzym, vol. 62, pp. 257-264, Mar. 2010.
 S. Sundar, J. Kundu and S. C. Kundu, “Biopolymeric nanoparticles,” Sci. Tech. Adv. Mater., vol. 11, pp. 1-13, Jan. 2010.
 J. Zhang, L. Liang, Z. Tian, L. Chen, and M. Subirade, “Preparation and in vitro evaluation of calcium-induced soy protein isolate nanoparticles and their formation mechanism study,” Food Chem., vol. 133, pp. 390-399, Jul. 2012.