摘要
将重组产几丁质酶C(Chi C)的菌体通过固定金属鳌合层析(IMAC),对目的蛋白进行初步纯化;然后采用透析法、凝胶过滤柱层析法和金属鳌合柱层析法对包涵体进行复性.结果表明,逐渐降低变性剂尿素浓度进行透析复性,酶液的酶活力为58.85μkat.L-1,比活为0.425 mkat.g-1,蛋白回收率为95.3%.使用SepHacry S-200作为复性凝胶介质复性,酶液的酶活力为30.37μkat.L-1,比活为0.620 mkat.g-1,蛋白回收率为72.9%.通过金属螯合层析法复性,酶液的酶活力为55.59μkat.L-1,比活为1.917 mkat.g-1,蛋白回收率为43.2%.
The recombinant proteins of the Recombinant Chitinase C Producing Escherichia coli are purified by passing through immobilized metal affinity chromatography（IMAC）,these inclusion bodies are renaturated by three Methods-membrane dialysis,gel filtration chromatography and IMAC.The results are ：（1） renaturated by gradually decreasing the urea concentration dialysis,the enzyme activity is 58.85 μkat·L-1,the specific activity is 0.425 mkat·g-1,and the protein recovery rate is 95.3%;（2）renaturated by SepHacry S-200 gel filtration chromatography,the enzyme activity is 30.37μkat·L-1,the specific activity is 0.620 mkat·g-1,and the protein recovery rate is 72.9%;（3）renaturated by IMAC,the enzyme activity is 55.59 μkat·L-1,the specific activity is 1.917 mkat·g-1,and the protein recovery rate is 43.2%.
出处
《河南师范大学学报（自然科学版）》
CAS
CSCD
北大核心
2011年第1期137-141,共5页
Journal of Henan Normal University(Natural Science Edition)
基金
福建省自然科学基金(C04010011)
