期刊论文

Kinetic modelling of thermal inactivation of His-Hep I and its application

中文

工业微生物

Industrial Microbiology

1001-6678

2014

44

2

46-51

10.3969/j.issn.1001-6678.2014.02.009

教育部博士点基金(No.20110093110008)； 江苏省自然科学基金(No.BK2011186)资助；

本校为其他机构

对组氨酸标签肝素酶I(His-Hep I, EC 4.2.2.7)的热失活机制进行了研究。针对短暂低温处理可以使热失活His-Hep I酶活部分恢复及添加二硫苏糖醇(DTT)使其热稳定性提高的现象,利用荧光探针法研究了失活过程His-Hep I构象变化,证明了该酶构象存在可逆转变行为。为进一步明晰His-Hep I的热失活机制,假设该酶热失活的主要途径包括去折叠及形成聚集体,并以此为基础建立模型进行拟合,模型与实验值吻合良好,表明假设的合理性。根据模型计算的活化能为E_r=100.217 kJ/mol、E_(ir)=7.857 kJ/mol和E_d=77.062 kJ/mol,此数据从一定程度上解释了冷处理为何能使His-Hep I部分恢复活性。进一步研究表明,任何能够...

The thermal inactivation found that lowering the temperature mechanism of his-tagged heparinase I （ His-Hep I, EC 4.2.2.7） was studied. It was could partially recover the enzymatic activity of the thermal inactivated enzyme and adding DTT could improve the enzyme stability. Therefore, fluorescence probe technique was adopted to monitor the conformation change of the enzyme during the thermal inactivation process. The results indicated that the process was reversible. Based on these phenomena, it was hypothesized that temperature induced prote...