An improved method of lipase preparation incorporating both solvent treatment and immobilization onto matrix |
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| Institution: | 1. Institute of Chemical and Engineering Sciences, 1 Pesek Road, Jurong Island, Singapore 627833, Singapore;2. Department of Chemical Science and Engineering, Kobe University, Nada-Ku, Kobe 657-8501, Japan;3. Graduate School of Science and Technology, Kobe University, Nada-Ku, Kobe 657-8501, Japan;1. Key Laboratory for Molecular Enzymology and Engineering, The Ministry of Education, Jilin University, Changchun 130012, China;2. National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China;3. School of Life Sciences, Jilin University, Changchun 130012, China;1. Instituto de Catálisis y Petroleoquímica (ICP), CSIC, Campus UAM-CSIC, Madrid, Spain;2. Food Biotechnology Division, Biotechnology Research Center (CRBt), Constantine, Algeria;3. Faculty of Nature and Life Sciences, Ibn Khalboun University, Tiaret, Algeria;4. Transformation and Food Product Elaboration Laboratory, Nutrition and Food Technology Institute (INATAA), Algeria;1. Universidade Federal do Rio de Janeiro, Faculdade de Farmácia, Departamento de Biotecnologia Farmacêutica, Centro de Ciências e Saúde, Bloco B, 1 andar (1 floor), Laboratório Multidisciplinar de Pesquisas em Biotecnologia, CEP 21941902, Rio de Janeiro, Brazil;2. Department of Biocatalysis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, 28049 Madrid, Spain;3. Departamento de Engenharia Química, Universidade Federal do Ceará, Campus do Pici, CEP 60455-760, Fortaleza, CE, Brazil;4. Programa de Pós-Graduação em Bioquímica, Departamento de Bioquímica, Brazil;5. Escuela de Química, Grupo de investigación en Bioquímica y Microbiología (GIBIM), Edificio Camilo Torres 210, Universidad Industrial de Santander, Bucaramanga, Colombia;1. Department of Applied Chemistry, School of Natural and Applied Sciences, Northwestern Polytechnical University, Xi’an 710072, China;2. MOE Key Laboratory of Material Physics and Chemistry under Extraordinary Conditions, Xi’an 710072, China;1. College of Pharmaceutical Science, Hebei University, Baoding 071002, China;2. Key Laboratory of Pharmaceutical Quality Control of Hebei Province, College of Pharmaceutical Science, Hebei University, Baoding 071002, China |
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| Abstract: | A simple and effective preparation of lipases for use in organic solvents is hereby proposed. Lipases in aqueous solution were treated with isopropanol, immediately followed by immobilization onto a commercially available macroporous resin CRBO2 (crosslinked polystyrene with N-methylglucamine as a functional group). The dual modification of lipases by (1) isopropanol treatment and (2) immobilization improved the activity and stability of lipases more significantly than either of the two treatments alone. The degree of lipase activation was dependent on isopropanol–buffer (v/v) ratio and the source of lipase used. Among the lipases tested, Rhizopus oryzae lipase was more significantly activated. The maximum specific activity of R. oryzae lipase after dual modification was 94.9 mmol h?1 g?1, which was, respectively, 3.3-, 2.5- and 1.5-fold of untreated free, untreated immobilized and treated free lipases. The conformations of the treated and untreated free lipases were investigated by circular dichroism (CD) measurement. Changes in the far- and near-UV CD spectra of lipase indicate that lipase activation is accompanied by changes in secondary and tertiary structures of lipases. The increase in negative molar elipticity at 222 nm suggests that the α-helical content of lipase increase after pretreatment. |
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