Covalent immobilization of lysozyme in silicone rubber modified by easy chemical grafting
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Research Letter
Covalent immobilization of lysozyme in silicone rubber modified by easy chemical grafting G. G. Flores-Rojas, Departamento de Química de Radiaciones y Radioquímica, Instituto de Ciencias Nucleares, Universidad Nacional Autónoma de México, Circuito Exterior, Ciudad Universitaria, México CDMX 04510, México; Nano Medical Engineering Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan F. López-Saucedo and E. Bucio, Departamento de Química de Radiaciones y Radioquímica, Instituto de Ciencias Nucleares, Universidad Nacional Autónoma de México, Circuito Exterior, Ciudad Universitaria, México CDMX 04510, México T. Isoshima, Nano Medical Engineering Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan Address all correspondence to G. G. Flores-Rojas at [email protected] and E. Bucio at [email protected] (Received 18 August 2017; accepted 2 October 2017)
Abstract Functionalization of silicone rubber films with lysozyme was achieved by grafting copolymerization and its chemical activation allowing the covalent immobilization of the enzyme. The new materials were characterized by means of Fourier-transform infrared spectroscopy, differential scanning calorimetry, thermogravimetric analysis, contact angle, atomic force microscopy, and mechanical properties of films. The enzymatic activity of films was studied by a suspension of lyophilized Micrococcus lysodeikticus. The activity test was inquired at different pH and temperatures, exhibiting enzymatic activity 20 °C above the free lysozyme, and at pH = 5 where the free lysozyme did not show activity.
Introduction The antimicrobial materials are the utmost importance in areas such as medical and food industry[1] and other areas where the bacteria growth is potential and require to inhibit it,[2] to preventing the bacterial attachment and improving the functionality, lifetime of devices. The medical area employs these materials in the prevention of clinic complications provoked by bacterial infections that sometimes even leading to death[3, 4] But in the case of food area, the antimicrobial materials are mainly required for the packaging and processing materials to hinder the accumulation of bacteria with a severe impact on the food quality, productivity, and processing efficiency.[5] The covalent immobilization of enzymes offers the possibility to provide to material with antimicrobial activity using the adequate enzyme[6] avoiding the growth of microorganism by passive antimicrobial activity. Preventing the formation of surface biofilms, and even though improving the enzymatic stability at a wide range of pH and temperature[7] due to the enzyme has its tertiary structure stabilized by the covalent bonds with the matrix. Lysozyme is the most commonly used enzyme to provide high antimicrobial activity[8] to materials,[7] using a moderate amount. Some methods applied for the enzyme immobilization has been by means of exchange reactions of thiol-disulfide,[9] formation of Schiff bases,[7] through the peptide[8] or diazo bonds[10] and alquilat
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