CotA漆酶的固定化及其对溴麝香草酚蓝脱色的效果  

Immobilization of CotA Laccase and Decoloring Bromothymol Blue

作　　者：邢丹[1] 马铭鸿 吴航伸 孙海琼[1] 王珏玉[1] 汪春蕾[1] 赵敏[1] 

机构地区：[1]东北林业大学,哈尔滨150040

出　　处：《东北林业大学学报》2017年第10期40-43,48,共5页Journal of Northeast Forestry University

基　　金：国家林业局"948"项目(2012-4-03);国家自然科学基金项目(51678120);黑龙江省博士后资助经费(LBH-Z11254)

摘　　要：将林地土壤中分离的枯草芽孢杆菌中克隆的CotA漆酶基因,在毕赤酵母菌中进行表达;以海藻酸钙为载体,采用包埋法固定毕赤酵母菌表达的CotA漆酶,固定化效率为32.2%,正交试验优化后的固定化效率比优化前提高了30.1%。优化了游离CotA漆酶及固定化CotA漆酶对溴麝香草酚蓝的脱色条件,在30min内,固定化CotA漆酶在70℃、pH为8.0时,游离CotA漆酶在60℃、pH为7.6时,对溴麝香草酚蓝溶液的脱色率最高,分别达到76.81%和67.12%,说明固定化CotA漆酶比游离CotA漆酶在较高温度和碱性条件下活性更高。CotA laccase gene from Bacillus subtilis isolated from forest soil was expressed in Pichia pastoris. The immobilization efficiency of CotA laccase expressed from Pichia pastoris was 32.2% with calcium alginate as carrier. The immobilization efficiency was increased by 30.1% through optimizing immobilization condition under orthogonal experiment. The highest decolorization rate of bromothymol blue by immobilized CotA laccase was 76.81% under 30 min, 70℃ and pH of 8.0. The highest decolorization rate of bromothymol blue by free laccase was 67.12% under 30 min, 60℃ and pH of 7.6. The results indicated that immobilized CotA laccase had higher enzyme activity than free CotA laccase under heat and alkali condition.

关 键 词：CotA漆酶 溴麝香草酚蓝 染料脱色 

分 类 号：Q554[生物学—生物化学] TQ61[化学工程—精细化工]