内生枯草芽孢杆菌产β-甘露聚糖酶的纯化研究  

Research on Purification of β - mannanase by Endophytic Bacillus subtilis

作　　者：张建新[1] 韩科芳[2] 赵杰[2] 曹金金[2] 邵亚萍[2] 赵念念[2] 

机构地区：[1]河南师范大学水产学院,河南新乡453007 [2]河南师范大学生命科学学院,河南新乡453007

出　　处：《生物技术》2013年第3期82-85,共4页Biotechnology

基　　金：河南省重点科技攻关项目(No.112102110118);河南省教育厅科技攻关项目(No.2010A180014)资助

摘　　要：方法:采用硫酸铵盐析、乙醇沉淀和聚乙二醇(PEG)/硫酸铵[(NH4)2SO4]双水相体系法对β-甘露聚糖酶进行提纯。目的:对一株内生枯草芽孢杆菌UD-20产生的β-甘露聚糖酶进行分离与纯化。结果:硫酸铵盐析与乙醇沉淀法提纯出酶的回收率分别为15%和7.5%,纯化倍数分别为1.968和0.977;双水相法提纯β-甘露聚糖酶在最优的体系为PEG2000 22%(m/m)、(NH4)2SO416%(m/m)、NaCl 2%(m/m)条件下,酶收率高达97.16%,萃取率为99.26%,纯化倍数为2.820。结论:硫酸铵盐析与乙醇沉淀法提纯出酶的回收率很低,而双水相法提纯β-甘露聚糖酶效果较好,可用于进一步的精提纯。Objective：Isolating and purifingβ- mannanase from an endophytic Bacillus subtilis strain UD- 20. Method：In the paper,β- mannanase was purified using ammonium sulfate salting -out, ethanol precipitation and polyethylene glycol （PEG） / ammonium sulfate [ （NH4）2 SO4 ] aqueous two - phase systems. Result：The recovery rates of the enzyme purified by （NH4 ）2 SO4 salting - out and ethanol precipitation were 15% and 7. 5%, respectively;The purificatioia multiples are 1. 968 and 0. 977, respectively. Under the optimal systems of PEG2000 22% （m/m）, （NH4）2SO, 16% （m/m） ,NaC1 2% （m/m） ,the recovery rate of the enzyme purified by aqueous two - phase systems was 97.16% ,the extraction yield was up to 99. 26% ,and the purification multiple was 2. 820. Conclusion：The recovery rates of the enzyme purified by （ NH4 ） 2 SO4 salting - out and ethanol precipitation are very low, but the aqueous two - phase systems was very well.

关 键 词：Β-甘露聚糖酶 枯草芽孢杆菌 双水相体系 盐析 酶收率 

分 类 号：TQ464[化学工程—制药化工]