木素过氧化物酶的逆胶束纯化研究

采用一种新型的逆胶束体系对木素过氧化物酶(LiP)进行了纯化.在此逆胶束体系中,以鼠李糖脂(RL,一种阴离子生物表面活性剂)作为表面活性剂,异辛烷和正己醇分别作为非极性有机溶剂和助表面活性剂(体积比为1:1),分别考察了RL浓度、振荡时间及水相的离子强度和pH值等因素对LiP的前萃取和后萃取过程的影响.结果表明,在前萃过程中,水相的最佳pH值为3.0,RL和KCl的最适浓度分别为2.75mmol/L和0.04mol/L,振荡时间以30min为最佳(200r/min);在后萃过程中,水相的最佳pH值为6.0,KCl的最适浓度为0.5mol/L.在上述最佳条件下,LiP的酶活回收率(AR)和纯化倍数(PF)分别高达93%, 2.9倍.电泳分析结果进一步证实LiP的纯化效果较佳.

The purification progress of lignin peroxidase by reversed mieelle system

A novel reversed micellar system had been developed to investigate the purification of lignin peroxidase(LiP).The anionic biosurfactant rhamnolipid(RL) was used to construct the reversed micellar system,while the isooctane and n-hexanol(1:1,V/V) were used as the apolar organic solvent and cosurfactant,respectively.In both the forward and backward extraction processes,the effects of RL concentration,shaking time,the pH value and ionic strength of water phase were studied.The results showed that the best condition for forward extraction was at 2.75mmol/L RL,0.04mol/L KCl,pH 3.0 and with the shaking time of 30min(200r/min).As regards backward extraction process,0.5mol/L KCl and pH 6.0 were suggested.In the above optimum conditions,the LiP activity recovery(AR) and purification fold(PF) were up to 93% and 2.9 times,respectively.The SDS-PAGE analysis further proved that the LiP was successfully purified.