双水相萃取分离重组毕赤酵母碱性木聚糖酶

Separation of alkaline xylanase from Pichia pastoris using aqueous two-phase extraction

利用双水相萃取技术对来源于重组毕赤酵母的碱性木聚糖酶进行了初步分离。研究了聚乙二醇(PEG)分子量、PEG浓度、盐种类及浓度、Na Cl加入量和p H等因素对萃取效果的影响。结果表明,在PEG 4000浓度为20%,Na2HPO4浓度为18%,Na Cl浓度为0,p H6.0的条件下,获得最佳萃取效果,其结果为:分配系数(K)8.51,酶的活力回收率(Y)87.2%,纯化倍数(PF)2.54。

Aqueous two-phase system was used to separate alkaline xylanase from recombinant Pichia pastoris.The effects of molecular weight of the PEG,PEG concentration,phase forming salts,salt concentration,neutral salt（Na Cl） concentration and p H on enzyme partitioning and purification were investigated. The results showed that PEG 4000 concentration 20%,disodium hydrogen phosphate concentration 18%,without Na Cl,under the condition of p H6.0 were the optimal conditions. And the best results were achieved as follows：partition coefficient（K） 8.51,enzyme activity recovery（Y） 87.2% and the purification factor（PF） 2.54.