title: Purification and in situ immobilization of lipase from of a mutant of Trichosporon laibacchii using aqueous two-phase systems.
abstract: The crude intracellular lipase (cell homogenate) from Trichosporon laibacchii was subjected to partial purification by aqueous two-phase system (ATPS) and then in situ immobilization by directly adding diatomites as carrier to the top PEG-rich phase of ATPS. A partition study of lipase in the ATPS formed by polyethylene glycol-potassium phosphate has been performed. The influence of system parameters such as molecular weight of PEG, system phase composition and system pH on the partitioning behaviour of lipase was evaluated. The ATPS consisting of PEG 4000 (12%) and potassium phosphate (K2HPO4, 13%) resulted in partition of lipase to the PEG-rich phase with partition coefficient 7.61, activity recovery 80.4%, and purification factor of 5.84 at pH of 7.0 and 2.0% NaCl. Moreover, the in situ immobilization of lipase in PEG phase resulted in a highest immobilized lipase activity of 1114.6 U g(-1). The above results show that this novel lipase immobilization procedure which couples ATPS extract and enzyme immobilization is cost-effective as well as time-saving. It could be potentially useful technique for the purification and immobilization of lipase.
authors.affiliation: Department of Pharmaceutics, Qingdao University of Science and Technology, 53 Zhengzhou Road, Qingdao, Shandong 266042, China.
authors.forename: Yuan-Yuan
authors.initials: YY
authors.lastname: Zhang
authors.forename: Jun-Hong
authors.initials: JH
authors.lastname: Liu
mesh_terms: D004800:Enzymes, Immobilized / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000302:isolation & purification*; D008970:Molecular Weight; D009154:Mutation / Q000235:genetics*; D044367:Phase Transition; D011092:Polyethylene Glycols; D014250:Trichosporon / Q000201:enzymology* / Q000235:genetics*; D014867:Water
chemical_list: D004800:Enzymes, Immobilized; D014867:Water; D011092:Polyethylene Glycols; D008049:Lipase