title: Isolation of a Aspergillus niger lipase from a solid culture medium with aqueous two-phase systems.
abstract: The aim of this work is to find the best conditions to isolate lipase from a solid culture medium of Aspergillus niger NRRL3 strains using aqueous two-phase systems formed with polyethylene glycol and potassium phosphate or polyethylene glycol and sodium citrate. We studied the partitioning of a commercial lyophilizate from A. niger. Also, the lipase enzymatic activity was studied in all the phases of the systems and the results indicate that citrate anion increases lipase activity. An analysis by fluorescence spectroscopy of the interaction between lipase and the bottom and top phases of the systems shows that the protein tryptophan-environments are modified by the presence of PEG and salts. Separation of the enzyme from the rest of the proteins that make up the lyophilized was achieved with good yield and separation factor by ATPS formed by PEG 1000/Pi at pH 7, PEG 2000/Ci at pH 5.2 and PEG 4000/Ci at pH 5.2. The above mentioned systems were used in order to isolate extracellular lipase from a strain of A. niger in submerged culture and solid culture. The best system for solid culture, with high purification factor (30.50), is the PEG 4000/Ci at pH 5.2. The enzyme was produced in a solid culture medium whose production is simple and recovered in a phase poor in polymer, bottom phase. An additional advantage is that the citrate produces less pollution than the phosphate. This methodology could be used as a first step for the isolation of the extracellular lipase from A. niger.
authors.affiliation: Physical-Chemistry Department, Faculty of Biochemical and Pharmaceutical Sciences, National University of Rosario, FonCyT, CIUNR and CONICET, Rosario, Argentina.
authors.forename: Analía
authors.initials: A
authors.lastname: Marini
authors.forename: Natalia
authors.initials: N
authors.lastname: Imelio
authors.forename: Guillermo
authors.initials: G
authors.lastname: Picó
authors.forename: Diana
authors.initials: D
authors.lastname: Romanini
authors.forename: Beatriz
authors.initials: B
authors.lastname: Farruggia
mesh_terms: D001234:Aspergillus niger / Q000201:enzymology*; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism; D019149:Bioreactors; D005591:Chemical Fractionation / Q000379:methods*; D002951:Citrates / Q000737:chemistry; D003470:Culture Media; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000302:isolation & purification* / Q000378:metabolism; D010710:Phosphates / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D002951:Citrates; D003470:Culture Media; D010710:Phosphates; D011092:Polyethylene Glycols; D008049:Lipase