Computer-aided design of bromelain and papain covalent immobilization

• Published in: Revista Colombiana de biotecnologia Vol. 16; no. 1; pp. 19 - 28
• Main Authors: Cutiño-Avila, Bessy, Gil Pradas, Dayrom, Aragón Abreu, Carlos, Fernández Marrero, Yuniel, Hernández de la Torre, Martha, Salas Sarduy, Emir, Chávez Planes, María De los Ángeles, Guisán Seijas, José Manuel, Díaz Brito, Joaquín, Del Monte-Martínez, Alberto
• Format: Journal Article
• Language: English; Spanish
• Published: Bogota Universidad Nacional de Colombia 01-01-2014; Instituto de Biotecnología, Universidad Nacional de Colombia
• Subjects: Biotechnology; BIOTECHNOLOGY & APPLIED MICROBIOLOGY; bromelain; bromelina; covalent immobilization; derivados inmovilizados; diseño racional; Enzymes; immobilized derivatives; inmovilización covalente; Ligands; papain; papaína; Proteases; rational design; rational design; diseño racional; derivados inmovilizados; papaína; covalent immobilization; immobilized derivatives; bromelina; inmovilización covalente; papain; bromelain
• ISSN: 0123-3475; 1909-8758; 1909-8758
• DOI: 10.15446/rev.colomb.biote.v16n1.44184

Enzymes as immobilized derivatives have been widely used in Food, Agrochemical, Pharmaceutical and Biotechnological industries. Protein immobilization is probably the most used technology to improve the operational stability of these molecules. Bromelain (Ananas comosus) and papain (Carica papaya) are cystein proteases extensively used as immobilized biocatalyst with several applications in therapeutics, racemic mixtures resolution, affinity chromatography and others industrial scenarios. The aim of this work was to optimize the covalent immobilization of bromelain and papain via rational design of immobilized derivatives strategy (RDID) and RDID1.0program. It was determined the maximum protein quantity to immobilize, the optimum immobilization pH (in terms of functional activity retention), and the most probable configuration of the immobilized derivative and the probabilities of multipoint covalent attachment was also predicted. As support material Glyoxyl-Sepharose CL 4B was used. The accuracy of RDID1.0 program´s prediction was demonstrated comparing with experimental results. Bromelain and papain immobilized derivatives showed desired characteristics for industrial biocatalysis, such as: elevate pH stability retaining 95% and 100% residual activity at pH 7.0 and 8.0, for bromelain and papain, respectively; high thermal stability at 30 °C retaining 90% residual activity for both immobilized enzymes; a catalytic configuration bonded by immobilization at optimal pH; and the ligand load achieved, ensures the minimization of diffusional restrictions.