Papain Immobilized Polyurethane Film as Antimicrobial Food Package

Authors: M. Cynthya, V. Prabhawathi, D. Mukesh

Abstract:

Food contamination occurs during post process handling. This leads to spoilage and growth of pathogenic microorganisms in the food, thereby reducing its shelf life or spreading of food borne diseases. Several methods are tried and one of which is use of antimicrobial packaging. Here, papain, a protease enzyme, is covalently immobilized with the help of glutarldehyde on polyurethane and used as a food wrap to protect food from microbial contamination. Covalent immobilization of papain was achieved at a pH of 7.4; temperature of 4°C; glutaraldehyde concentration of 0.5%; incubation time of 24h; and 50mg of papain. The formation of -C=Nobserved in the Fourier transform infrared spectrum confirmed the immobilization of the enzyme on the polymer. Immobilized enzyme retained higher activity than the native free enzyme. The modified polyurethane showed better reduction of Staphylococcus aureus biofilm than bare polymer film (eight folds reduction in live colonies, two times reduction in protein and 6 times reduction in carbohydrates). The efficacy of this was studied by wrapping it over S. aureus contaminated cottage cheese (paneer) and cheese and stored at a temperature of 4°C for 7days. The modified film reduced the bacterial contamination by eight folds when compared to the bare film. FTIR also indicated reduction in lipids, sugars and proteins in the biofilm.

Keywords: Cheese, Papain, polyurethane, Staphylococcus aureus.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1097287

References:

[1] Mangalassary. S, Antimicrobial Food Packaging to Enhance Food Safety: Current Developments and Future Challenges, J. Food Process Technol, vol.3, pp.1-2, 2012.
[2] Calzolai. L, Gilliland. D, Rossi. F, Measuring nanoparticles size distribution in food and consumer products: a review, Food. Addit. Contam. Part A, vol. 29, pp.1183-1193, 2012.
[3] Veluchamy.P, Sivakumar. P.M, Doble.M, Immobilization of Subtilisin on Polycaprolactam for Antimicrobial Food Packaging Applications, J. Agric. Food Chem, vol.59, pp.10869-10878, 2011.
[4] Sarker. S.D, Nahar. L, Kumarasamy. Y, Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals, Methods, vol.43, pp.321-324, 2007.
[5] Fernandez-Lafuente.R, Rossell.C.M, Rodriguez.V, Santana.C, Soler. G, Bastida. A, Guisan. A. M, Preparation of activated supports containing low pK amino groups. A new tool for protein immobilization via the carboxyl coupling method, Enzyme Microb. Technol, vol.15, pp.546– 550, 1993.
[6] Lopez-Gallego. F, Betancor. L, Hidalgo. A, Mateo. C, Guisan. J. M, Fernandez-Lafuente. R, Optimization of an industrial biocatalyst of glutaryl acylase: Stabilization of the enzyme by multipoint covalent attachment onto new amino-epoxy sepabeads. J. Biotechnol, vol. 111, pp.219–227, 2004.
[7] Sivakumar. P. M, Prabhawathi. V, Doble. M, Chalcones as an effective antibiofoulant against marine isolated microorganisms, Colloids Surf. B, vol. 81, pp. 439–446, 2010.
[8] Lowry. O.H, Rosenbrough. N.J, Farr. A.L, Randall. R. J, Protein measurement with the folin phenol reagent, J. Biol. Chem, vol. 193, pp.265-275, 1951.
[9] Lowry. O.H, Rosenbrough. N.J, Farr. A.L, Randall. R. J, Protein measurement with the folin phenol reagent, J. Biol. Chem, vol. 193, pp.265-275, 1951.
[10] Besse. N. G, Audinet. N, Beaufort. A, Colin. P, Cornu. M, Lombard. B, A contribution to the improvement of Listeria monocytogenes enumeration in cold-smoked salmon, Int J Food Microbiol, vol. 91, pp.119-127. 2004.
[11] Prabhawathi. V, Boobalan. T, Sivakumar. P.M, Doble. M, Antibiofilm Properties of Interfacially Active Lipase Immobilized Porous Polycaprolactam Prepared by LB Technique, Plos One, vol. 5, pp.1-11, 2014.
[12] Pavithra, D., Doble, M., Biofilm formation, bacterial adhesion and host response on polymeric implants issues and prevention. Biomed. Mater.3 (3),4003-11,2008.