US EPA

6575020 

Journal Article 

Gene cloning and characterization of thiourocanate hydratase from Burkholderia sp. HME13 

Muramatsu, H; Miyaoku, H; Kurita, S; Matsuo, H; Kashiwagi, T; Kim, CS; Hayashi, M; Yamamoto, H; Kato, SI; Nagata, S; , 

2020 

Yes 

Journal of Biochemistry
ISSN: 0021-924X
EISSN: 1756-2651 

OXFORD UNIV PRESS 

OXFORD 

167 

3 

333-341 

English 

31725161 

10.1093/jb/mvz098 

WOS:000526615500013 

http://://WOS:000526615500013
Exit
 

A novel enzyme, thiourocanate hydratase, which catalyses the conversion of thiourocanic acid to 3-(5-oxo-2-thioxoimidazolidin-4-yl) propionic acid, was isolated from the ergothioneine-utilizing strain, Burkholderia sp. HME13. When the HME13 cells were cultured in medium containing ergothioneine as the sole nitrogen source, thiourocanate-metabolizing activity was detected in the crude extract from the cells. However, activity was not detected in the crude extract from HME13 cells that were cultured in Luria-Bertani medium. The gene encoding thiourocanate hydratase was cloned and expressed in Escherichia coli, and the recombinant enzyme was purified to homogeneity. The enzyme showed maximum activity at pH 7.5 and 55°C and was stable between pH 5.0 and 10.5, and at temperatures up to 45°C. The Km and Vmax values of thiourocanate hydratase towards thiourocanic acid were 30 μM and 7.1 μmol/min/mg, respectively. The enzyme was strongly inhibited by CuCl2 and HgCl2. The amino acid sequence of the enzyme showed 46% identity to urocanase from Pseudomonas putida, but thiourocanate hydratase had no urocanase activity. 

ergothioneine; thiourocanic acid; thiourocanate hydratase; urocanase; Burkholdria; ergothioneine; biosynthesis; alignment; mushrooms; Biochemistry & Molecular Biology