Platform design for extraction and isolation of Bromelain: Complex formation and precipitation with carrageenan |
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| Affiliation: | 1. CBQF-Centro de Biotecnologia e Química Fina-Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa/Porto, Rua Arquiteto Lobão Vital Apartado 2511, 4202-401, Porto, Portugal;2. IPROBYQ-Institute of Biotechnological and Chemistry Processes, Faculty of Biochemical and Pharmaceutical Sciences and CONICET, National University of Rosario, Suipacha 570, (S2002RLK) Rosario, Argentina;3. INL-International Iberian Nanotechnology Laboratory, 4710-330, Braga, Portugal;4. Centro de Engenharia Biológica, Universidade do Minho, Campus Gualtar, 4710-057, Braga, Portugal;1. Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Republic of Korea;2. R&D Center, Daesang Corporation, Gyeonggi-Do 467-813, Republic of Korea;3. Department of Food Science and Technology, Department of Interdisciplinary Bio IT Materials, Seoul National University of Science and Technology, Seoul 139-743, Republic of Korea;4. Center for Food and Bioconvergence, Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-742, Republic of Korea;1. Université de Liège, Laboratoire d’Enzymologie et Repliement des Protéines, Centre d’Ingénierie des Protéines, Liège, Belgium;2. Université Libre de Bruxelles, Faculty of Medicine, Protein Chemistry Unit, Campus Erasme (CP 609), 808 route de Lennik, 1070 Brussels, Belgium;1. School of Food and Biological Engineering, Jiangsu University, Zhenjiang, Jiangsu, 212013, China;2. School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, Jiangsu, 212013, China;3. Henan Key Laboratory of Water Pollution Control and Rehabilitation Technology, Henan University of Urban Construction, Pingdingshan, Henan, 467036, China;4. School of Pharmaceutical Science and Technology, Suzhou Chien-shiung Institute of Technology, Taicang, 215400, China |
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| Abstract: | The main objective of this work was to investigate for the first time the molecular mechanism of complex formation between bromelain (a positively charged enzyme) and carrageenan (a natural strong polyelectrolyte, negatively charged) using spectroscopy techniques and thermodynamic approaches. The Bromelain-Carrageenan complex showed a maximal non-solubility at pH around 5.1. The solubility was dependent on pH and ionic strength of the medium. To re-dissolve the formed complex, the pH was changed and 500 mM of NaCl was added to the initial solution, proving the columbic mechanism for the formation of non-soluble complex. The formation of the carrageenan-bromelain complex increased in 8 °C the enzyme thermal stability, while its biological activity was not modified. The amount of total enzyme recovered in solution after precipitation with around 0.08% w/v of carrageenan was 85–90%. |
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| Keywords: | Protein complex Enzyme mechanism Carrageenan Polyelectrolytes Bromelain |
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