Cloning, expression and characterization of catechol 1,2-dioxygenase from Burkholderia cepacia
| Autor: | Nguyen Duc Huy, Duong Duc Hoang Sinh, Tran Vu Ngoc Thi, Nguyen Hoang Loc, Nguyen Hoang Tue, Kazuhide Kimbara, Masaki Shintani, Le Thi Ha Thanh |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
Cloning 0303 health sciences Catechol biology Chemistry 16S ribosomal RNA biology.organism_classification 01 natural sciences Applied Microbiology and Biotechnology Microbiology High-performance liquid chromatography Molecular biology Enzyme assay law.invention 03 medical and health sciences chemistry.chemical_compound Burkholderia law 010608 biotechnology Recombinant DNA biology.protein Catechol 1 2-dioxygenase 030304 developmental biology |
| Zdroj: | The Journal of General and Applied Microbiology. 66:188-194 |
| ISSN: | 1349-8037 0022-1260 |
| Popis: | The present study reports on the cloning, expression and characterization of catechol 1,2-dioxygenase (CAT) of bacterial strains isolated from dioxin-contaminated soils in Vietnam. Two isolated bacterial strains DF2 and DF4 were identified as Burkholderia cepacia based on their 16S rRNA sequences. Their genes coding CAT was amplified with a specific pair of primers. Recombinant CAT (rCAT) was expressed in E. coli M15 cells and its activity was confirmed by the detection of cis,cis-muconic acid, a product from catechol, by high-performance liquid chromatography (HPLC) analysis. The rCAT of DF4 had an optimal pH and temperature of 7 and 30°C, respectively. Metal ions, such as Zn2+ and Mn2+, and surfactants, such as SDS, Tween 20 and Triton X100, strongly inhibited enzyme activity, while K+ slightly increased the activity. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|