One-step three-phase partitioning of lipoxygenase from defatted soy flours
CAI Yan1, TIAN Dan1, YAN Xin1, LI Baiyu3, LI Yujie1, YU Lijuan2*, WU Jinming1*
1(School of Chemistry and Chemical Engineering, Nantong University,Nantong 226019,China) 2(School of Life Science,Nantong University,Nantong 226019,China) 3(School of Pharmaceutical and Environmental Engineering,Nantong Vocational University,Nantong 226007,China)
Abstract: Three phase partitioning (TPP) for partial purification of lipoxygenases (LOXs) in one step is reported for the first time. The influences of solvent, temperature, pH and dosage of ammonium sulfate on purification fold and activity recovery were investigated. The optimized conditions were as follows: the volume ratio of crude enzyme to tert butyl alcohol was 1∶2.5, the saturation of (NH4)2SO4 was 60%, the pH was 8, the temperature was 20 ℃, and the reaction time was 60 min. LOXs was purified by 8.2 folds and 78.2% activity was retained. SDS-PAGE analysis of partially purified enzyme revealed a molecular weight of around 95 kDa. TPP was proved to be an effective technique for the partial purification of LOXs from defatted soy flours.
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