类芽孢杆菌产β-甘露聚糖酶的分离与纯化  

Separation and purification of β-mannanase from Paenibacillus sp.

作　　者：张建新[1] 穆广亚 陈琳[1] 李玲玲[1] 聂国兴[1] 

机构地区：[1]河南师范大学水产学院,河南新乡453007

出　　处：《中国酿造》2016年第10期68-71,共4页China Brewing

基　　金：河南省重点科技攻关(132102310293;142102210453);河南省教育厅科学技术研究重点项目(14B180023);河南师范大学国家级科研项目培育基金(2013PL10)

摘　　要：类芽孢杆菌(Paenibacillus sp.)CH-3是一株高产β-甘露聚糖酶的菌株。本研究采用乙醇沉淀法、硫酸铵盐析法、聚乙二醇(PEG)/硫酸铵[(NH4)2SO4]双水相体系对CH-3菌株产生的β-甘露聚糖酶进行分离与纯化,以选出纯化率高且残余酶活力高的方法。结果发现,乙醇沉淀法和硫酸铵盐析法纯化效果较差,聚乙二醇(PEG)/硫酸铵[(NH4)2SO4]双水相体系的纯化效果较好,PEG浓度为20%(m/m)时,上相测得的总酶活力最高,可以达到221 U/m L。在(NH4)2SO4浓度为18%(m/m)时,上相测得总酶活力最高,可以达到264 U/m L,同时酶萃取率达最大值为86.2%。因此,最优的双水相体系是20%(m/m)PEG和浓度为18%(m/m)(NH4)2SO4。Paenibacillus sp. CH-3 is a selective breeding bacterial strain of β-mannase with high yield. In order to choose the method with high purification rate and high residual enzyme activity,β-mannanase is purified by ammonium sulfate salting-out, ethanol precipitation and polyethylene glycol （PEG）/ammonium sulfate [（NH4）2SO4] aqueous two-phase systems. The result shows that ethanol precipitation method and ammonium sulfate salting-out method have poor purification effect, while polyethylene glycol/ammonium sulfate aqueous two-phase systems has better purification effect. When the PEG concentration is 20%（m/m）, the total supematant enzyme activity is the highest （221 U/mL）. Meanwhile the extraction yield is up to 89.6%. When the [（NH4）2SO4] concentration is 18%（m/m）, the total supernatant enzyme activity is the highest, the extraction yield is up to 86.2%. Therefore, the concentration of 20%（m/m） PEG and the concentration of 18%（m/m） （NI-h）2SO4 aqueous two-phase system could be the best.

关 键 词：Β-甘露聚糖酶 提取纯化 双水相体系 

分 类 号：TQ464[化学工程—制药化工]