来源于红球菌胆固醇氧化酶ChOG的异源表达、纯化及催化反应结构分析

Subjects: Biology >> Bioengineering submitted time 2017-06-01 Cooperative journals: 《中国生物工程杂志》

张玉富 王建文 李松涛 朱张亮 路福平 毛淑红 秦慧民

Abstract： Cholesterol oxidase is a key enzyme during cholesterol metabolism. It could be potential used to detect the cholesterol level in serum in clinical trials. The cholesterol oxidase from Rhodococcus ruber (ChOG) was transformed into BL21(DE3) and Rosetta(DE3) for protein expression, and was induced at different conditions. The results showed that ChOG was overexpressed at 16 ℃ and 0.1 mmol/L IPTG. The activity reached 8.033 U/mg after purification with Ni-NTA superflow. The product cholest-5-en-3-one was characterized by TLC and HPLC, respectively. The structural analysis showed that Glu406, together with Arg408 and Glu261 played an important role during the dehydrogenation of cholesterol C3-OH, proton transfer, and isomerization.

 Hits 19920  Downloads 1830  Comment 0