|
Characterization of a recombinant arylsulfatase enzyme from glucosinolate-metabolizing human gut bacterium Escherichia coli VL8 |
|---|---|
| รหัสดีโอไอ | |
| Creator | Vijitra Luang-In |
| Title | Characterization of a recombinant arylsulfatase enzyme from glucosinolate-metabolizing human gut bacterium Escherichia coli VL8 |
| Contributor | John T. Rossiter, Abdulhadi A. Albaser |
| Publisher | Asia-Pacific Journal of Science and Technology |
| Publication Year | 2566 |
| Journal Title | Asia-Pacific Journal of Science and Technology |
| Journal Vol. | 28 |
| Journal No. | 2 |
| Page no. | 9 |
| Keyword | Desulfo-glucosinolate, Escherichia coli VL8, p-Nitrocatechol sulfate, Recombinant enzyme, Sulfatase |
| URL Website | https://www.tci-thaijo.org/index.php/APST |
| Website title | https://so01.tci-thaijo.org/index.php/APST/article/view/252493 |
| ISSN | 2539-6293 |
| Abstract | Bacterial arylsulfatases can hydrolyze organosulfur compounds. Thus, the objective of this research project was to characterize a recombinant arylsulfatase (ARS) from Escherichia coli VL8, a human gut bacterium able to produce nitrile from desulfo-glucosinolates. The Ars gene (Accession no.: LC685335.1) with a length of 1,494 bp corresponding to ARS enzyme with a length of 497 amino acids was cloned from E. coli VL8 and expressed in E. coli BL21 (DE3) for 16 h at 25?C in lysogeny broth (LB) by induction of isopropyl ?-D-1-thiogalactopyranoside (0.5 mM). The recombinant ARS enzyme (57 kDa) was partially purified using Ni2+ affinity column chromatography. This recombinant ARS enzyme was able to desulfate synthetic p-nitrocatechol sulfate (pNCS) substrate to produce p-nitrocatechol as an indicator of ARS activity, with optimal conditions at 30?C and pH 6.0, respectively. The ARS enzyme displayed a Michaelis-Menten kinetic constant (Km) of 1.09 mM and Maximum reaction velocity (Vmax) of 25.1 U/mg for pNCS. ARS enzyme activity toward pNCS was not enhanced by any metal ions, while activity was inhibited by Ca2+, Fe2+, NaHSO4 and Na2SO4. |
