title: Potential application of aqueous two-phase systems and three-phase partitioning for the recovery of superoxide dismutase from a clarified homogenate of Kluyveromyces marxianus.
abstract: Superoxide dismutase (SOD; EC 1.15.1.1) is an antioxidant enzyme that represents the primary cellular defense against superoxide radicals and has interesting applications in the medical and cosmetic industries. In the present work, the partition behavior of SOD in aqueous two-phase systems (ATPS) (using a standard solution and a complex extract from Kluyveromyces marxianus as sample) was characterized on different types of ATPS (polymer-polymer, polymer-salt, alcohol-salt, and ionic liquid (IL)-salt). The systems composed of PEG 3350-potassium phosphate, 45% TLL, 0.5 M NaCl (315 U/mg, 87% recovery, and 15.1-fold purification) and t-butanol-20% ammonium sulfate (205.8 U/mg, 80% recovery and 9.8-fold purification), coupled with a subsequent 100 kDa ultrafiltration stage, allowed the design of a prototype process for the recovery and partial purification of the product of interest. The findings reported herein demonstrate the potential of PEG-salt ATPS for the potential recovery of SOD.
authors.affiliation: Centro de Biotecnología FEMSA, Tecnológico de Monterrey, Monterrey, NL, 64849, México.
authors.forename: Jesús
authors.initials: J
authors.lastname: Simental-Martínez
authors.forename: Marco
authors.initials: M
authors.lastname: Rito-Palomares
authors.forename: Jorge
authors.initials: J
authors.lastname: Benavides
mesh_terms: D018533:Biomass; D002853:Chromatography, Liquid / Q000379:methods*; D000431:Ethanol / Q000737:chemistry; D052578:Ionic Liquids / Q000737:chemistry; D007716:Kluyveromyces / Q000201:enzymology*; D010710:Phosphates / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D017680:Potassium Compounds / Q000737:chemistry; D012965:Sodium Chloride / Q000737:chemistry; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000302:isolation & purification*; D014462:Ultrafiltration / Q000379:methods*
chemical_list: D052578:Ionic Liquids; D010710:Phosphates; D017680:Potassium Compounds; D000431:Ethanol; D011092:Polyethylene Glycols; D012965:Sodium Chloride; C013216:potassium phosphate; D013482:Superoxide Dismutase
keywords: Kluyveromyces marxianus; aqueous two-phase systems; enzyme recovery; superoxide dismutase; three-phase partitioning