FTIR-ATR characterization of free Rhizomucor meihei lipase (RML), Lipozyme RM IM and chitosan-immobilized RML |
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| Affiliation: | 1. INTEC, UNL-CONICET, Güemes 3450, Santa Fe 3000, Argentina;2. INQUISUR, UNS, Avenida Alem 1253, 8000 Bahía Blanca, Argentina;3. PLAPIQUI-UNS-CONICET, Camino La Carrindanga Km 7, CC 717, 8000 Bahía Blanca, Argentina;1. Centro Regional de Estudios en Alimentos Saludables, Valparaíso, Chile;2. Laboratorio de Síntesis de Péptidos e Inmunología Molecular, Pontificia Universidad Católica de Valparaíso, Núcleo de Biotecnología Curauma, Valparaíso, Chile;3. Grupo Ciencia de los Materiales, Instituto de Química, Facultad de Ciencias Exactas y Naturales, Universidad de Antioquia UdeA, Calle 70 No. 52-21, Medellín, Colombia;4. Grupo de Biocatálisis, Escuela de Ingeniería Bioquímica, Facultad de Ingeniería, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile;1. Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, Bandar Sunway, 47500 Subang Jaya, Selangor, Malaysia;2. Monash-Industry Palm Oil Education and Research Platform (MIPO), Monash University Malaysia, Jalan Lagoon Selatan, Bandar Sunway, 47500 Subang Jaya, Selangor, Malaysia;1. Cukurova University, Imamoglu Vocational School, Adana, Turkey;2. Cukurova University, Sciences & Letters Faculty, Chemistry Department, 01330, Adana, Turkey;3. Cukurova University, Ceyhan Engineering Faculty, Chemical Engineering Department, Adana, Turkey;4. Departamento De Biocatalisis, ICP-CSIC, C/Marie Crue 2, Campus UAM-CSIC, Cantoblanco, 28049, Madrid, Spain;5. Center of Excellence in Bionanoscience Research, Member of The External Scientific Advisory Board, King Abdulaziz University, Jeddah, Saudi Arabia;1. Department of Chemical Engineering, Faculty of Engineering, University of Guilan, P. O. Box 41635-3756, Rasht, Iran;2. Department of Chemistry, Payame Noor Unversity, P. O. Box 19395-3697, Tehran, Iran;1. Energy and Environment Unit, Engineering and Processing Research Division, Malaysian Palm Oil Board, 6, Persiaran Institusi, Bandar Baru Bangi, 43000 Kajang, Selangor, Malaysia;2. Department of Chemical and Environmental Engineering, Faculty of Engineering, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia;3. Institute of Plantation Studies, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia;4. Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia |
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| Abstract: | The synthesis and characterization of biocatalysts based on lipase from Rhizomucor miehei (RML) immobilized on chitosan-based supports were investigated. The enzyme was immobilized on chitosan following two strategies: (i) physical adsorption; and (ii) covalent bonding using glutaraldehyde. The content of enzyme bound in the supports, as precipitable protein, was analyzed using UV/visible methods. FTIR-ATR spectroscopy was employed to characterize the prepared biocatalysts, as well the native enzyme and a commercial biocatalyst Lipozyme RM IM, used as reference materials. Analysis of the amide I′ signal allowed to follow the changes in the secondary structure of the enzyme after binding to the support and its thermal stability. The hydrolysis of ethyl stearate monitored in situ by FTIR-ATR was used as a test reaction. Results showed that RML was bound to Chit and Glut–Chit with minor changes in its secondary structure, thermal stability and enzymatic activity in a selected reaction test. |
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