聚乙二醇/硫酸铵双水相体系萃取大豆脂肪氧合酶的研究

Extraction of soybean lipoxygenase by PEG / (NH4) 2SO4 aqueous Two- phase systems

摘要: 采用聚乙二醇(PEG)/硫酸铵(NH4)2SO4双水相体系对大豆中脂肪氧合酶进行分离萃取研究;通过综合考察酶分配系数、相比和回收率,探讨了(NH4)2SO4、PEG2000、Na Cl浓度以及p H对脂肪氧合酶萃取的影响,并通过正交实验进一步优化实验条件,结果表明在单一因素下的最佳工艺条件为:(NH4)2SO415%(wt%)、PEG2000 13%(wt%)、Na Cl 1%(wt%)、p H5.8;正交实验优选出的最佳工艺条件为:(NH4)2SO417%(wt%)、PEG2000 13%(wt%)、p H5.2,可获得酶的分配系数最高可达9.710,萃取率为87.6%。

Abstract: Separation of lipoxygenase directly from nonfat soybean in aqueous two- phase systems ( ATPS) of polyethylene glycol / ammonium sulfate was studied. The effects of polyethylene glycol 2000 ( PEG2000) concentration, ( NH4) 2SO4concentration, Na Cl concentration, and p H value on the extraction of soybean lipoxygenase were investigated in terms of partition coefficients, phase volume of both enzyme activity and recovery.Orthogonal experiment was used to further analyze and optimize the separation of lipoxygenase. The results indicated that the optimum separation conditions of single factor conditions were PEG2000 13%, ( NH4) 2SO415%, Na Cl 1% and p H5.8. Maximum distribution coefficient and yield of lipoxygenase was 9.710 and87.6% under the optimum separation conditions of PEG2000 13%, ( NH4) 2SO417% and p H5.2 obtained by orthogonal experiment.