分枝杆菌的代谢研究。III. 麻风分枝杆菌中三羧酸循环关键酶的证明。

Metabolic studies on mycobacteria. III. Demonstration of key enzymes of TCA cycle in M. leprae.

作者信息

Katoch V M, Sharma V D, Kannan K B, Datta A K, Shivannavar C T, Bharadwaj V P

机构信息

Dept. of Microbiology, Central JALMA Institute for Leprosy, Taj Ganj.

出版信息

Indian J Lepr. 1987 Apr-Jun;59(2):152-7.

PMID:3309085

Abstract

Cell free extracts of armadillo derived M. leprae, M. phlei, M. smegmatis and normal armadillo liver were analysed for the two key enzymes of TCA cycle. Aconitase activity was assayed in the presence of inhibitor fluorocitrate and it was observed that cell free extracts from cultivable mycobacteria as well as aramadillo derived M. leprae had this enzyme activity and 66-82% of this activity was inhibited by 0.1 mM fluorocitrate. 74% of M. leprae derived enzyme activity was inhibited by fluorocitrate in contrast with armadillo derived enzyme which was only 29% inhibited by fluorocitrate. PAGE separation of cell free extracts and staining for Isocitrate dehydrogenase (ICD) activity showed that an additional bond of ICD activity was demonstrable in the cell free extracts of armadillo derived M. leprae and this was NADP dependent. The mobility (ef) of this band of activity was in the same range as ICD from cultivable mycobacteria and much lower than ICD from normal armadillo liver. From this study and from the previously reported work, it is concluded that like other mycobacteria TCA cycle is operative in M. leprae.

摘要

对犰狳源麻风分枝杆菌、草分枝杆菌、耻垢分枝杆菌以及正常犰狳肝脏的无细胞提取物进行了三羧酸循环两种关键酶的分析。在存在抑制剂氟柠檬酸盐的情况下测定乌头酸酶活性，结果观察到可培养分枝杆菌以及犰狳源麻风分枝杆菌的无细胞提取物具有这种酶活性，且0.1 mM氟柠檬酸盐可抑制该活性的66 - 82%。与犰狳源酶仅29%被氟柠檬酸盐抑制形成对比的是，犰狳源麻风分枝杆菌的酶活性74%被氟柠檬酸盐抑制。对无细胞提取物进行聚丙烯酰胺凝胶电泳分离并对异柠檬酸脱氢酶（ICD）活性进行染色，结果显示在犰狳源麻风分枝杆菌的无细胞提取物中可证明存在一条额外的ICD活性条带，且该条带依赖于NADP。这条活性条带的迁移率（ef）与可培养分枝杆菌的ICD处于相同范围，远低于正常犰狳肝脏的ICD。从本研究以及先前报道的工作可以得出结论，与其他分枝杆菌一样，三羧酸循环在麻风分枝杆菌中也有作用。