Evaluating aqueous two-phase systems for Yarrowia lipolytica extracellular lipase purification |
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| Institution: | 1. Escola de Química, Universidade Federal do Rio de Janeiro, Av. Athos da Silveira Ramos, 149 – CT, Bl. E, Ilha do Fundão, 21941-909, Rio de Janeiro, Brazil;2. Laboratório de Nanotecnologia Biofuncional, Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Av. Carlos Chagas Filho, 373 – Cidade Universitária, 21941-170, Rio de Janeiro, Brazil;3. Departamento de Tecnologia Rural e Animal – DTRA, Universidade Estadual do Sudoeste da Bahia, Rodovia BR 415, Km 03, S/N, 45700-000, Itapetinga, Bahia, Brazil;4. Departamento de Ciências Exatas e Tecnológicas DCET, Universidade Estadual de Santa Cruz, Rod. Jorge Amado, km 16 – Salobrinho, 45662-900, Ilhéus, Bahia, Brazil;1. Division of Microbial Resources, Chemical, Biological and Agricultural Pluridisciplinary Research Center (CPQBA), Campinas State University (UNICAMP), SP, Brazil;2. Biotechnology Interunit Post-graduation Program USP/IPT/Butantã, São Paulo, SP, Brazil;3. Department of Biochemical and Pharmaceutical Technology, School of Pharmaceutical Sciences, University of São Paulo, Brazil;4. Department of Biochemistry and Microbiology, University of São Paulo State (UNESP/Rio Claro), SP, Brazil;1. Department of Chemical and Environmental Engineering, Faculty of Engineering, University of Nottingham Malaysia Campus, Jalan Broga, 43500 Semenyih, Selangor Darul Ehsan, Malaysia;2. CICECO – Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal;1. Faculty of Food Science and Technology, University Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia;2. Institute of Technology, Middle Technical University, 29008 Alzafaranya, Baghdad, Iraq;3. Halal Products Research Institute, University Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia;1. Universidade Tiradentes, Av. Murilo Dantas 300, Farolândia, 49032-490 Aracaju-SE, Brazil;2. ITP, Instituto de Tecnologia e Pesquisa, Av. Murilo Dantas, 300-Prédio do ITP, Farolândia, 49032-490 Aracaju-SE, Brazil;3. Departamento de Química, CICECO, Universidade de Aveiro, 3810-193 Aveiro, Portugal;1. Department of Biophysics, Biosciences Institute (IB), UFRGS, CEP 91501-970, Porto Alegre, RS, Brazil;2. Graduate Program in Cellular and Molecular Biology ? Center for Biotechnology, Universidade Federal do Rio Grande do Sul (UFRGS), CEP 91501-970, Porto Alegre, RS, Brazil;3. Instituto do Cérebro (InsCer) and Graduate Program in Medicine and Health Sciences, Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), CEP 90610-000, Porto Alegre, RS, Brazil;4. Graduate Program in Cellular and Molecular Biology Applied to Health, Genetics Applied Toxicology, Universidade Luterana do Brasil (ULBRA), CEP 92425-900, Canoas, RS, Brazil;5. Department of Molecular Biology and Biotechnology, IB, UFRGS, CEP 91501-970, Porto Alegre, RS, Brazil;6. Department of Biology, University of Toronto Mississauga, Mississauga, Ontario, Canada |
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| Abstract: | In this study an aqueous two-phase system (ATPS) composed of polyethylene glycol (PEG) and potassium phosphate was tested for the purification of lipase from Yarrowia lipolytica IMUFRJ 50682. Ultrafiltration and precipitation with acetone and kaolin were also used as traditional comparison methods Ultrafiltration was a good method with a purification factor of 6.55, but protease was also purified in this extract. For the precipitation with acetone and kaolin lower values of lipase and protease activity were found in relation to the original crude enzyme extract. Under the best conditions of ATPS (pH 6 and 4 °C), the purification fold was greater than 40 and selectivity was almost 500. Lipase was recovered in the salty phase which makes it easier to purify it. The optimum pH and temperature ranges for purified lipase with this system was 6–7 and 35–40 °C, respectively. Lipase thermostability was increased in relation to crude extract after the purification with the PEG/phosphate buffer system for temperatures lower than 50 °C. All enzyme extracts showed good stability to a wide pH range. Y. lipolytca lipase was successfully purified by using ATPS in a single downstream processing step and presented good process characteristics after this treatment. |
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| Keywords: | Lipases Aqueous two-phase system Ultrafiltration Enzyme stability Purification |
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