Document Type : Original research

Authors

1 Q.C and Analyzer of Samin Laboratory, Urmia, Iran

2 Department of food science and technology, Faculty of agriculture, Urmia University

3 R&D department of Shirin Asal food industry, Tabtiz, Iran

Abstract

The aim of this research was to compare the effect of free and immobilized Lysozyme on the physicochemical and antimicrobial properties of sodium caseinate (SC) based active films. Lysozyme was immobilized onto bacterial cellulose nanofiber (BCNF) and decreasing of its activity was approved after immobilization. Free and immobilized enzymes were incorporated into SC films at the concentrations of 0.5 and 1 mg.100ml-1 and the films were characterized. Addition of BCNF and lysozyme diminished significantly (p˂0.05) the moisture absorption and water vapor permeability of SC films and immobilized enzyme had higher effect than free enzyme on decreasing of these parameters. The tensile strength and Young’s modulus were increased and elongation to break was decreased by incorporation of BCNF. An adverse effect was observed for lysozyme addition but the effect of immobilized enzyme on the weakening of tensile properties was lower than free lysozyme. According to X-ray diffraction (XRD) results, the crystallinity of films increased by incorporation of BCNF and lysozyme. However, the immobilization reduced the crystalline regions of BCNF. Antimicrobial activity of lysozyme increased after immobilization and SC films containing immobilized enzyme exhibited considerable activity against Gram-positive bacteria S. aureus, L. monocytogenes. Gram-negative bacteria E. coli, Y. enterocolitica, mold of A. niger and yeast of S. cerevisiae.

Keywords

Abdollahi, M., Alboofetileh, M., Behrooz, R., Rezaei, M., & Miraki, R. (2013). Reducing water sensitivity of alginate bio-nanocomposite film using cellulose nanoparticles. International Journal of Biological Macromolecules, 54, 166-173.
Abdul Hanid, N., UzirWahit, M., Guo, Q., Mahmoodian, S., & Soheilmoghaddam, M. (2014). Development of regenerated cellulose/halloysitesnanocomposites via ionic liquids. Carbohydrate Polymers, 99, 91–97.
Abouhmad, A., Dishisha, T., Amin, M.A., Hatti-Kual, R. (2017). Immobilization to positively charged cellulose nanocrystals enhances the antibacterial activity and stability of hen egg white and T4 lysozyme. Biomacromolecules, 18(5) 1600–1608.
Achaby, E.M.,  Kassab, Z.,  Barakat, A., & Aboulkas, A. (2018). Alfa fibers as viable sustainable source for cellulose nanocrystals extraction: Application for improving the tensile properties of biopolymer nanocomposite films. Industrial Crops & Products, 112, 499–510
Aliheidari, N., Fazaeli, M., Ahmadi, R., Ghasemlou, M., & Emam-Djomeh, Z. (2013). Comparative evaluation on fatty acid and Matricaria recutita essential oil incorporated into casein-based film. International Journal of Biological Macromolecules, 56, 69–75.
Almasi, H., Ghanbarzadeh, B., Dehghannya, J., Entezami, A. A., & Khosrowshahi Asl, A. (2015a). Novel nanocomposites based on fatty acid modified cellulose nanofibers/poly (lactic acid): Morphological and physical properties. Food Packaging and Shelf Life, 5, 21-31.
Almasi, H., Ghanbarzadeh, B., Dehghannya, J., Pirsa, S. & Zandi, M. (2015b). Heterogeneous modification of softwoods cellulose nanofibers with oleic acid: effect of reaction time and oleic acid concentration. Fibers and Polymers, 16, 1715-1722.
Angles, M.N., & Dufresne, A. (2000). Plasticized starch/tunicin whiskers nanocomposites. 1. Structural analysis. Macromolecules, 33(22), 8344-8353.
Arrieta, M.P., Peltzer, M.A., López , J., Garrigós, M.C., Valente, Artur.J.M., & Jiménez, A. (2014). Functional properties of sodium and calcium caseinate antimicrobial active films containing carvacrol. Journal of Food Engineering, 121, 94-101.
ASTM. (2005). Standard test methods for water vapor transmission of material. E96-05. Annual book of ASTM, Philadelphia, PA: American Society for Testing and Materials.
ASTM. (2010). Standard test methods for tensile properties of thin plastic sheeting. D882-10. Annual book of ASTM, Philadelphia, PA: American Society for testing and Materials.
Atarés, L., Bonilla, J., & Chiralt, A. (2010). Characterization of sodium caseinate-based edible films incorporated with cinnamon or ginger essential oils. Journal of Food Engineering, 100, 678-687.
Aulin, C., Ahola, S., Josefsson, P., Nishino, T., Hirose, Y., Österberg, M., & Wagberg, L., (2009). Nanoscale cellulose films with different crystallinities and mesostructures: their surface properties and interaction with water. Langmuir, 25(13), 7675–7685.
Azeredo, H., Mattoso, L.H.C., Avena-Bustillos, R.J., Munford, M.L., Wood, D., & McHugh, T.H. (2010). Nanocellulose reinforced chitosan composite films as affected by nanofiller loading and plasticizer content. Journal of Food Science, 75, N1–N7.
Bayazidi, P., Almasi, H. & Khosrowshahi Asl, A. (2018). Immobilization of lysozyme on bacterial cellulose nanofibers: characteristics, antimicrobial activity and morphological properties. International Journal of Biological Macromolecules, 107, 2544-2551.
Benucci, I., Liburdi, K., Cacciotti, I., Lombardelli, C., Zappino, M., Nanni, F., & Esti, M. (2018). Chitosan/clay nanocomposite films as supports for enzyme immobilization: An innovative green approach for winemaking applications. Food Hydrocolloids, 74, 124-131.
Cao, W-T., Chen, F-F., Zhu, Y-J., Zhang, Y-G., Jiang, Y-Y., Ma, M-G., & Chen, F. (2018). Binary strengthening and toughening of MXene/cellulose nanofiber composite paper with nacre-inspired structure and superior electromagnetic interference shielding properties. American Chemical Society, 12, 4583−4593.
Cappannella, E., Benucci, I., Lombardelli, C., Liburdi, K., Bavaro, T., & Esti, M. (2016). Immobilized lysozyme for the continuous lysis of lactic bacteria in wine: Bench-scale fluidized-bed reactor study. Food Chemistry, 210, 49–55.
Carrillo, W., Garcíaruiz, A., Recio, I., & Morenoarribas, M.V. (2014). Antibacterial activity of hen egg white lysozyme modified by heat and enzymatic treatments against oenological lactic acid bacteria and acetic acid bacteria. Food Protection, 77(10), 1732–1739.
Chaichi, M., Hashemi, M., Badii, F., & Mohammadi, A. (2017). Preparation and characterization of a novel bionanocomposite edible film based on pectin and crystalline nanocellulose. Carbohydrate Polymers, 157, 167-175.
Chen, Q., Liu, D., Wu, C., Yao, K., Li, Z., Shi, N., Wen, F., & Gates, I.D. (2018). Co-immobilization of cellulase and lysozyme on amino-functionalized magnetic nanoparticles. Bioresource Technology, 263, 317-324.
Chevalier, E., Assezat, G., Prochazka, F., & Oulahal, N. (2018a). Development and characterization of a novel edible extruded sheet based on different casein sources and influence of the glycerol concentration. Food Hydrocolloids, 75, 182-191.
Chevalier, E., Chaabani, A., Assezat, G., & Prochazka, F. (2018b). Casein/wax blend extrusion for production of edible films as carriers of potassium sorbate: A comparative study of waxes and potassium sorbate effect. Food Packaging and Shelf Life, 16, 41–50.
Coma, V. (2008) Bioactive packaging technologies for extended shelf life of meat-based products. Journal of Meat Science, 78, 90–103.
Fu, J., Li, D., Li, G., Huang, F. & Wei, Q. (2015). Carboxymethyl cellulose assisted immobilization of silver nanoparticles onto cellulose nanofibers for the detection of catechol. Journal of Electroanalytical Chemistry, 738, 92-99.
Flores-Rojas, Guadalupe G., Pino-Ramos, Victor H., López-Saucedo, F., Concheiro, A., Alvarez-Lorenzo, C., & Bucio, E. (2017). Improved covalent immobilization of lysozyme on silicon rubber films grafted with poly(ethylene glycol dimethacrylate-coglycidyl methacrylate). European Polymer Journal, 95, 27–40.
Ghanbarzadeh, B., & Almasi, H. (2011). Physical properties of edible emulsified films based on carboxymethyl cellulose and oleic acid. International Journal of Biological Macromolecules, 48(1), 44-49.
Ghanbarzadeh, B., Oleyaei, S. A., & Almasi, H. (2015). Nanostructured materials utilized in biopolymer-based plastics for food packaging applications. Critical Reviews in Food Science and Nutrition, 55(12), 1699-1723.
Graebin, N., Andrades, D., Bonin, M., Rodrigues, R., & Ayub, M. (2016). Dextransucrase immobilized on activated-chitosan particles as a novel biocatalyst. Journal of Molecular Catalysis B: Enzymatic, 133, S143-S149.
Hamou, B K., Kaddami, H., Dufresne, A., Boufi, S., Magnin, A., & Erchiqui, F. (2018). Impact of TEMPO-oxidization strength on the properties of cellulose nanofibril reinforced polyvinyl acetate nanocomposites. Carbohydrate Polymers, 181, 1061–1070.
Hanušová, K., Vápenka, L., Dobiáš, J., & Mišková, L. (2013). Development of antimicrobial packaging materials with immobilized glucose oxidase and lysozyme.  Central Europea Journal of Chemistry, 11(7), 1066-1078.
Homaei, AA. Sariri, R., Vianello, F., & Stevanato, R. (2013). Enzyme immobilization: an update. Journal of Chemical Biology, 6, 185-205.
Kalia, S., Boufi, S., Celli, A., & Kango, S. (2014). Nanofibrillated cellulose: surface modification and potential applications. Colloid Polymer Science, 292, 5-31.
Kim, J., Grate, W.J., & Wang, P. (2008). Nanobiocatalysis and its potential applications. Trends in Biotechnology, 26, 639-646.
Kuorwel, K., Marlene J. C., Kees, S., Joseph, M., & Stephen W.B. (2012). Evaluation antifungal activity of antimicrobial agents on cheddar cheese. Packaging Technology & Science, 14428.
Kuorwel, K.K., Cran, M.J., Sonneveld, K., Miltz, J. & Bigger, SW. (2014). Evaluation of antifungal activity of antimicrobial agents on Cheddar cheese. Packaging Technology & Science, 27, 49-58.
Li, J., Si, Y., Zhao, C., He, J., Sun, G., & Huang, Y. (2017). Spontaneous and efficient adsorption of lysozyme from aqueous solutions by naturally polyanion gel beads. Materials Science and Engineering C: Materials for Biological Applications, 76, 130-138.
Lian, Z.X., Ma, Z.S., Wei, J., & Liu, H. (2012). Preparation and characterization of immobilized lysozyme and evaluation of its application in edible coatings. Process Biochemistry, 47, 201-208.
Liu, Y., Edwards, J.V., Prevost, N., Huang, Y., & Chen. Y,J. (2018). Physico- and bio-activities of nanoscale regenerated cellulose nonwoven immobilized with lysozyme. Materials Science & Engineering: C, 91, 389-394.
Nakimbugwe, D., Masschalck, B., Deckers, D., Callewaert, L., Atanassova, M., Aertsen, A,. & Michiels C.W. (2005). Purification of Ivy, a lysozyme inhibitor from Escherichia coli, and characterization of its specificity for various lysozymes. Enzyme and Microbial Technology, 37(2), 205-211.
Park, J., Kim, M., Park, H-S., Jang, A., Min, J,. & Kim, Y-H. (2013). Immobilization of lysozyme-CLEA onto electrospun chitosan nanofiber for effective antibacterial applications. International Journal of Biological Macromolecules, 54, 37-43. 
Pereda, M., Amica, G., Racz, I,. & Marcovich, N.E. (2011a). Structure and properties of nanocomposite films based on sodium caseinate and nanocellulose fibers. Journal of Food Engineering, 103(1), 76–83.
Pereda, M., Amica, G., Rácz, I., & Marcovich, N.E. (2011b). Preparation and characterization of sodium caseinate films reinforced with cellulose derivatives. Carbohydrate Polymers, 86(2), 1014-1021.
Picchio, M., Monti, G., Gugliotta, L., Minari, R., & Alvarez Igarzabal, C. (2018). Casein films crosslinked by tannic acid for food packaging applications. Food Hydrocolloids, 84, 424-434.
Pourjavaher, S., Almasi, H., Meshkini, S., Pirsa, S., & Parandi, E. (2017). Development of a colorimetric pH indicator based on bacterial cellulose nanofibers and red cabbage (Brassica oleraceae) extract. Carbohydrate Polymers, 156, 193–201.
Safarik, I., Pospiskova, K., Baldikova, E,. & Safarikova, M. (2016). Magnetically responsive biological materials and their applications. Advanced Materials Letters, 7, 254-261.
Salgado, P. R., Molina Ortiz, S. E., Petruccelli, S., & Mauri, A. N. (2010). Biodegradable                 sunflower protein films naturally activated with antioxidant compounds. Food Hydrocolloids, 24, 525–533.
Schou, M., Longares, A., Montesinos-Herrero, C., Monahan, F. J., O’Riordan, D., & O’sullivan, M. (2005). Properties of edible sodium caseinate films and their application as food wrapping. LWT-Food Science and Technology, 38(6), 605-610.
 Shabanpour, B., Kazemi, M., S., Ojagh, M., & Pourashouri, P. (2016). Bacterial cellulose nanofibers as reinforce in edible fish myofibrillar protein nanocomposite films. International Journal of Biological Macromolecules, 117, 742-751.
Shankar, S., Teng, X., & Rhim, J.W. (2014). Properties and characterization of agar/CuNP bionanocomposite films prepared with different copper salts and reducing agents. Carbohydrate Polymers, 114, 21517-21524.
Shankar, S., Teng, X., Li, G., & Rhim, J. W. (2015). Preparation, characterization, and antimicrobial activity of gelatin/ZnO nanocomposite films. Food Hydrocolloids, 45, 264-271.
Sirisha, V.L., Jain, A. & Jain, A. (2016). Enzyme Immobilization: An Overview on Methods, Support Material, and Applications of Immobilized Enzymes. Advances in Food and Nutrition Research, 79, 179-211.
Sun, J., Yendluri, R., Liu, K., Guo, Y., Lvov, Y., & Yan, X. (2016). Enzyme-immobilized clay nanotube–chitosan membranes with sustainable biocatalytic activities. Physical Chemistry Chemical Physics, 19, 562-567.
Uddin, khan M.A., Orelma, H., Mohammadi, P., Borghei, M., Laine, J., Linder, M., & Rojas, Orland J. (2017). Retention of lysozyme activity by physical immobilization in nanocellulose aerogels and antibacterial effects. Cellulose, 24, 2837-2848.
Wagh, Y. R., Pushpadass, Heartwin A., Magdaline Eljeeva Emerald, F., & Surendra Nath, B. (2013). Preparation and characterization of milk protein films and their application for packaging of Cheddar cheese. Journal of Food Science & Technology, 51, 3767-3775.
Wang, D., Lv, R., Ma, X., Zou, M., Wang, W., Yan, L., Ding, T., Ye, X., & Liu, D. (2018). Lysozyme immobilization on the calcium alginate film under sonication: Development of an antimicrobial film. Food Hydrocolloids, 83, 1-8.
Zhang, Y., Liu, Y., Li, R., Ren, X., & Huang, T.S. (2018). Preparation and characterization of antimicrobial films based on nanocrystalline cellulose. Applied Polymer Science, In Press. DOI: 10.1002/app.47101.
Zhang, Y., Nypelo¨, T., Salas, C., Arboleda, J., Hoeger, IC. & Rojas, OJ. (2013). Cellulose nanofibrils: from strong materials to bioactive surfaces. Renewable Materials, 1(3), 195–211.