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Purification and characterization of a collagenase from Penicillium sp. UCP 1286 by polyethylene glycol-phosphate aqueous two-phase system.
de Albuquerque Wanderley, Maria Carolina; Wanderley Duarte Neto, José Manoel; Campos Albuquerque, Wendell Wagner; de Araújo Viana Marques, Daniela; de Albuquerque Lima, Carolina; da Cruz Silvério, Sara Isabel; de Lima Filho, José Luiz; Couto Teixeira, José António; Porto, Ana Lúcia Figueiredo.
Afiliação
  • de Albuquerque Wanderley MC; Laboratory of Immunopathology Keizo Asami (LIKA), Federal University of Pernambuco, Av. Prof. Moraes Lins do Rego, s/n, 50670-901, Recife, PE, Brazil.
  • Wanderley Duarte Neto JM; Laboratory of Immunopathology Keizo Asami (LIKA), Federal University of Pernambuco, Av. Prof. Moraes Lins do Rego, s/n, 50670-901, Recife, PE, Brazil.
  • Campos Albuquerque WW; Department of Morphology and Animal Physiology, Federal Rural University of Pernambuco, Av. Dom Manoel de Medeiros, s/n, 52171-900, Recife, PE, Brazil.
  • de Araújo Viana Marques D; Serra Talhada Campus, University of Pernambuco (UPE), Av. Afonso Magalhães, s/n, Serra Talhada, PE, Brazil.
  • de Albuquerque Lima C; Faculty of Science, Education and Technology of Garanhuns, University of Pernambuco-UPE, Av. Capitão Pedro Rodrigues, n° 105, Garanhuns, PE, Brazil.
  • da Cruz Silvério SI; Centre of Biological Engineering, University of Minho, Campus Gualtar, 4710-057, Braga, Portugal.
  • de Lima Filho JL; Laboratory of Immunopathology Keizo Asami (LIKA), Federal University of Pernambuco, Av. Prof. Moraes Lins do Rego, s/n, 50670-901, Recife, PE, Brazil.
  • Couto Teixeira JA; Centre of Biological Engineering, University of Minho, Campus Gualtar, 4710-057, Braga, Portugal.
  • Porto ALF; Department of Morphology and Animal Physiology, Federal Rural University of Pernambuco, Av. Dom Manoel de Medeiros, s/n, 52171-900, Recife, PE, Brazil. Electronic address: analuporto@yahoo.com.br.
Protein Expr Purif ; 133: 8-14, 2017 05.
Article em En | MEDLINE | ID: mdl-28242427
Collagenases are proteolytic enzymes capable of degrading both native and denatured collagen, reported to be applied in industrial, medical and biotechnological sectors. Liquid-liquid extraction using aqueous two-phase system (ATPS) is one of the most promising bioseparation techniques, which can substitute difficult solid-liquid separation processes, offering many advantages over conventional methods including low-processing time, low-cost material and low-energy consumption. The collagenase produced by Penicillium sp. UCP 1286 showed a stronger affinity for the bottom salt-rich phase, where the highest levels of collagenolytic activity were observed at the center point runs, using 15.0% (w/w) PEG 3350 g/mol and 12.5% (w/w) phosphate salt at pH 7.0 and concentration. The enzyme was characterized by thermal stability, pH tolerance and effect of inhibitors, showing optimal collagenolytic activity at 37 °C and pH 9.0 and proved to be a serine protease. ATPS showed high efficiency in the collagenase purification, confirmed by a single band in SDS/PAGE, and can in fact be applied as a quick and inexpensive alternative method.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Penicillium / Fosfatos / Polietilenoglicóis / Proteínas Fúngicas / Colagenases Idioma: En Revista: Protein Expr Purif Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2017 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos
Texto completo
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Penicillium / Fosfatos / Polietilenoglicóis / Proteínas Fúngicas / Colagenases Idioma: En Revista: Protein Expr Purif Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2017 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos