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Partitioning of lactate dehydrogenase from bovine heart crude extract by polyethylene glycol-citrate aqueous two-phase systems |
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| Authors: | Rosângela Ferreira Frade de Araújo Tatiana Souza Porto Danyelly Bruneska Gondim Martins Rosa Fireman DutraAna Lúcia Figueiredo Porto José Luiz de Lima Filho |
| Institution: | a Departamento de Bioquímica, Universidade Federal de Pernambuco, Recife, PE, Brazil b Laboratório de Imunopatologia Keizo Asami, LIKA, Universidade Federal de Pernambuco, Recife, PE, Brazil c Departamento de Morfologia e Fisiologia Animal, Universidade Federal Rural de Pernambuco, PE, Brazil d Departamento de Patologia, Universidade de Pernambuco, Recife, PE, Brazil |
| Abstract: | Aqueous two-phase systems (ATPS) composed of polyethylene glycol (PEG)-citrate have been used for enzyme partitioning studies. The behavior of lactate dehydrogenase (LDH) from bovine heart crude extract was analyzed using a two-level factorial design in which the PEG molar mass and concentration, the citrate concentration were selected as independent variables, while the purification factor, the partition coefficient (K) and the activity yield were selected as responses. The statistical analysis revealed the effect of PEG molar mass on K. LDH exhibited a better partitioning toward PEG-rich phase and the highest K value (1079.81) was obtained with 42% (w/w) PEG 400 and 7.5% (w/w) citrate concentration. PEG molar mass also influenced the purification factor of the enzyme in the top phase. Possibly these ATPS remove inhibitors present in the extract affording higher enzyme yield. |
| Keywords: | Aqueous two-phase system Partitioning Lactate dehydrogenase Polyethylene glycol Citrate |
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