基因工程菌1020耐热木聚糖酶的纯化  

Purification of Thermo-stable Xylanase from Bioengineered Bacterium 1020

作　　者：孙雷[1] 朱孝霖[1] 李环[1] 姚忠[1] 韦萍[1] 

机构地区：[1]南京工业大学制药与生命科学学院,江苏南京210009

出　　处：《生物技术》2005年第5期51-53,共3页Biotechnology

摘　　要：目的:提纯基因工程菌1020耐热木聚糖酶。方法:超声破碎细胞,而后经过热变性处理和Ni-NTA亲和层析分离纯化。结果:根据pET System Manual的方法分析木聚糖酶主要分布在可溶性细胞质中;超声破壁功率400W,破碎15min时效果最佳;粗酶液经过70℃,热变性30min处理后,纯化倍数达到4.9;采用Ni-NTA Sephrose F.F一步层析可以得到电泳纯的木聚糖酶,纯化倍数13.4,得率29.4%。结论:热变性处理是一种有效的提纯手段,合理的条件可以使纯化倍数提高;Ni-NTA亲和层析是纯化含His标签的目的酶的特异高效纯化方法。Objective：Thermo- stable xylanase from bioengineered bacterium 1020 was purified to homogeneity.Methods： Ultrasonic was used to disrupt cells. After heat treatment, the crude enzyme solution was loaded on Ni- NTA Sepharose F. F to purify. Results： The xylanse was expressed in soluble cytoplasic fraction.That ultrasonic powder was 400W, and disrupted ceils about 15min is best for this experlment.The purified factor can reach 4.9 by heat treat treatment at 70℃ for 30min. Only Ni - NTA affinity chromatography can make the xylanase to homogeneity. The purified factor is 13.4 and recovery is 29.4%. Conclusions： Heat treatment is a good purification method for thermostable xylanase; Ni - NTA affinity chromatography is a biospecial chromatography for xylanase fused with His - tag.

关 键 词：耐热木聚糖酶 纯化 热变性 Ni—NTA亲和层析 

分 类 号：Q556.2[生物学—生物化学]