title: Extraction protease expressed by Penicillium fellutanum from the Brazilian savanna using poly(ethylene glycol)/sodium polyacrylate/NaCl aqueous two-phase system.
abstract: The partitioning of protease expressed by Penicillium fellutanum from the Brazilian savanna in a novel inexpensive and stable aqueous two-phase system (ATPS) composed of poly(ethylene glycol) (PEG) and sodium polyacrylate (NaPA) was studied in this work using factorial design. The ATPS is formed by mixing both polymers with a salt (NaCl) and fermented broth of P. fellutanum. The effects of molar mass (2,000, 4,000, and 6,000 g ⋅ mol(-1)) and concentration (6, 8, and 10 wt%) of PEG and that of NaPA concentration (6, 8, and 10 wt%) on protease partitioning (K) at 25 °C were studied. A two-level factorial design (2(3)) was implemented. The effect of Na2 SO4 concentration (5, 10, and 15 wt%) on the reextraction of the enzyme was also analyzed. The partition coefficient K ranged from 77.51 to 1.21, indicating the versatility of the method. The reextraction was achieved with the addition of 5% Na2 SO4 , allowing the partitioning of the protease to the upper phase, whereas total proteins were directed to the bottom phase. The results of partitioning using the PEG/NaPA/NaCl system and that of the subsequent reextraction with Na2 SO4 suggest that this method can be used to purify proteases from fermented broth of P. fellutanum.
authors.affiliation: Department of Pharmaceutical Sciences, School of Health Sciences, University of Brasilia, Brasilia, DF, Brazil.
authors.forename: Kleber V G
authors.initials: KV
authors.lastname: Barros
authors.affiliation: Laboratory of Enzymology, Department of Cellular Biology, University of Brasilia, Brasilia, DF, Brazil.
authors.forename: Paula M
authors.initials: PM
authors.lastname: Souza
authors.affiliation: Department of Pharmaceutical Sciences, School of Health Sciences, University of Brasilia, Brasilia, DF, Brazil.
authors.forename: Samuel L
authors.initials: SL
authors.lastname: Cardoso
authors.affiliation: Department of Pharmaceutical Sciences, School of Health Sciences, University of Brasilia, Brasilia, DF, Brazil.
authors.forename: Leonardo L
authors.initials: LL
authors.lastname: Borges
authors.affiliation: Department of Biochemical and Pharmaceutical Technology, University of São Paulo, São Paulo, SP, Brazil.
authors.forename: Edivaldo X F
authors.initials: EX
authors.lastname: Filho
authors.affiliation: Laboratory of Enzymology, Department of Cellular Biology, University of Brasilia, Brasilia, DF, Brazil.
authors.forename: Adalberto P
authors.initials: AP
authors.lastname: Junior
authors.affiliation: Department of Pharmaceutical Sciences, School of Health Sciences, University of Brasilia, Brasilia, DF, Brazil.
authors.forename: Pérola O
authors.initials: PO
authors.lastname: Magalhães
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry*; D005591:Chemical Fractionation / Q000379:methods*; D015870:Gene Expression; D065948:Grassland; D010407:Penicillium / Q000235:genetics*; D010447:Peptide Hydrolases / Q000235:genetics / Q000302:isolation & purification*; D011092:Polyethylene Glycols / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry*; D014867:Water / Q000737:chemistry
chemical_list: D000180:Acrylic Resins; D014867:Water; D011092:Polyethylene Glycols; D012965:Sodium Chloride; C006903:carbopol 940; D010447:Peptide Hydrolases
keywords: factorial design; fermented broth; poly(acrylic acid); poly(ethylene glycol); protease; purification