title: Purification and characterization of a collagenase from Penicillium sp. UCP 1286 by polyethylene glycol-phosphate aqueous two-phase system.
abstract: 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.
authors.affiliation: Laboratory of Immunopathology Keizo Asami (LIKA), Federal University of Pernambuco, Av. Prof. Moraes Lins do Rego, s/n, 50670-901, Recife, PE, Brazil.
authors.forename: Maria Carolina
authors.initials: MC
authors.lastname: de Albuquerque Wanderley
authors.affiliation: Laboratory of Immunopathology Keizo Asami (LIKA), Federal University of Pernambuco, Av. Prof. Moraes Lins do Rego, s/n, 50670-901, Recife, PE, Brazil.
authors.forename: José Manoel
authors.initials: JM
authors.lastname: Wanderley Duarte Neto
authors.affiliation: Department of Morphology and Animal Physiology, Federal Rural University of Pernambuco, Av. Dom Manoel de Medeiros, s/n, 52171-900, Recife, PE, Brazil.
authors.forename: Wendell Wagner
authors.initials: WW
authors.lastname: Campos Albuquerque
authors.affiliation: Serra Talhada Campus, University of Pernambuco (UPE), Av. Afonso Magalhães, s/n, Serra Talhada, PE, Brazil.
authors.forename: Daniela
authors.initials: D
authors.lastname: de Araújo Viana Marques
authors.affiliation: Faculty of Science, Education and Technology of Garanhuns, University of Pernambuco-UPE, Av. Capitão Pedro Rodrigues, n° 105, Garanhuns, PE, Brazil.
authors.forename: Carolina
authors.initials: C
authors.lastname: de Albuquerque Lima
authors.affiliation: Centre of Biological Engineering, University of Minho, Campus Gualtar, 4710-057, Braga, Portugal.
authors.forename: Sara Isabel
authors.initials: SI
authors.lastname: da Cruz Silvério
authors.affiliation: Laboratory of Immunopathology Keizo Asami (LIKA), Federal University of Pernambuco, Av. Prof. Moraes Lins do Rego, s/n, 50670-901, Recife, PE, Brazil.
authors.forename: José Luiz
authors.initials: JL
authors.lastname: de Lima Filho
authors.affiliation: Centre of Biological Engineering, University of Minho, Campus Gualtar, 4710-057, Braga, Portugal.
authors.forename: José António
authors.initials: JA
authors.lastname: Couto Teixeira
authors.affiliation: 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.
authors.forename: Ana Lúcia Figueiredo
authors.initials: ALF
authors.lastname: Porto
mesh_terms: D017364:Collagenases / Q000737:chemistry / Q000302:isolation & purification*; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification*; D010407:Penicillium / Q000201:enzymology*; D010710:Phosphates / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*
chemical_list: D005656:Fungal Proteins; D010710:Phosphates; D011092:Polyethylene Glycols; D017364:Collagenases
keywords: ATPS; Collagen; Collagenolytic enzyme; Purification