Expression, purification, and activity of ActhiS, a thiazole biosynthesis enzyme from Acremonium chrysogenum
- 作者: Song Z.1, Pan J.1, Xie L.1, Gong G.1, Han S.1, Zhang W.1, Hu Y.1
-
隶属关系:
- China State Institute of Pharmaceutical Industry
- 期: 卷 82, 编号 7 (2017)
- 页面: 852-860
- 栏目: Article
- URL: https://journals.rcsi.science/0006-2979/article/view/151442
- DOI: https://doi.org/10.1134/S0006297917070112
- ID: 151442
如何引用文章
详细
Thiamine pyrophosphate is an essential coenzyme in all organisms. Its biosynthesis involves independent syntheses of the precursors, pyrimidine and thiazole, which are then coupled. In our previous study with overexpressed and silent mutants of ActhiS (thiazole biosynthesis enzyme from Acremonium chrysogenum), we found that the enzyme level correlated with intracellular thiamine content in A. chrysogenum. However, the exact structure and function of ActhiS remain unclear. In this study, the enzyme-bound ligand was characterized as the ADP adduct of 5-(2-hydroxyethyl)-4-methylthia-zole-2-carboxylic acid (ADT) using HPLC and 1H NMR. The ligand-free ActhiS expressed in M9 minimal medium catalyzed conversion of NAD+ and glycine to ADT in the presence of iron. Furthermore, the C217 residue was identified as the sulfur donor for the thiazole moiety. These observations confirm that ActhiS is a thiazole biosynthesis enzyme in A. chrysogenum, and it serves as a sulfur source for the thiazole moiety.
作者简介
Zhihui Song
China State Institute of Pharmaceutical Industry
Email: bebydou@hotmail.com
中国, Shanghai, 201203
Jie Pan
China State Institute of Pharmaceutical Industry
Email: bebydou@hotmail.com
中国, Shanghai, 201203
Liping Xie
China State Institute of Pharmaceutical Industry
Email: bebydou@hotmail.com
中国, Shanghai, 201203
Guihua Gong
China State Institute of Pharmaceutical Industry
Email: bebydou@hotmail.com
中国, Shanghai, 201203
Shu Han
China State Institute of Pharmaceutical Industry
Email: bebydou@hotmail.com
中国, Shanghai, 201203
Wei Zhang
China State Institute of Pharmaceutical Industry
Email: bebydou@hotmail.com
中国, Shanghai, 201203
Youjia Hu
China State Institute of Pharmaceutical Industry
编辑信件的主要联系方式.
Email: bebydou@hotmail.com
中国, Shanghai, 201203