大连工业大学学报2025，Vol.44Issue(2)：109-114,6.DOI:10.19670/j.cnki.dlgydxxb.2025.0206
N-脱甲基酶电子转移受体蛋白CdnD及其Rieske型[2Fe-2S]结构域的功能表征
Functional characterization of N-demethylase electrons transfer receptor protein CdnD and its Rieske type[2Fe-2S]domain
摘要
Abstract
A gene named cdnD from caffeine degrading bacterium Paraburkholderia caffeinelytica CF1 is on the caffeine degradation related gene cluster and corresponding to the N-demethylase electron transfer receptor protein.Its function was determined by heterologous expression,purification and enzyme activity The function of CdnD was determined by heterologous expression,purification and enzyme activities.It has been found that there is an additional Rieske type[2Fe-2S]domain in CdnD compared to other oxidoreductases in the same family.To investigate the role of the domain in the N-demethylation process,the Rieske type[2Fe-2S]domain of CdnD was knocked out and the mutant strain CdnDΔD1 was constructed.Its effects on the activities of N1-demethylase CdnA and N7-demethylase CdnC were studied respectively.The results showed that the N1 methyl group of the substrate could be removed by CdnDΔD1 combined with CdnA,but the N7 methyl group of the substrate could not be removed by CdnDΔD1 combined with CdnC and CdeE.Therefore,it has been proved that the electrons could be transported by the Rieske type[2Fe-2S]domain of CdnD in the catalytic process of N7-demethylation.关键词
咖啡因/N-脱甲基酶/Rieske型[2Fe-2S]/电子转移受体蛋白Key words
caffeine/N-demethylase/Rieske type[2Fe-2S]/electrons transfer receptor protein分类
生物学引用本文复制引用
李会佳,燕云,孟亦轲,高子晴..N-脱甲基酶电子转移受体蛋白CdnD及其Rieske型[2Fe-2S]结构域的功能表征[J].大连工业大学学报,2025,44(2):109-114,6.基金项目
辽宁省民族药功效成分开发与应用重点实验室开放课题(2021JH13/10200038). （2021JH13/10200038）
