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Characterization of a recombinant ?-glucosidase of GH3 familyfrom glucosinolate-metabolizing human gut bacteriumEnterococcus casseliflavus CP1 for nitrile production |
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| รหัสดีโอไอ | |
| Creator | 1. Vijitra Luang-In 2. Abdulhadi Ali Albaser 3. John T. Rossiter |
| Title | Characterization of a recombinant ?-glucosidase of GH3 familyfrom glucosinolate-metabolizing human gut bacteriumEnterococcus casseliflavus CP1 for nitrile production |
| Publisher | Research and Development Office, Prince of Songkla University |
| Publication Year | 2563 |
| Journal Title | Songklanakarin Journal of Science and Technology |
| Journal Vol. | 42 |
| Journal No. | 3 |
| Page no. | 549-556 |
| Keyword | ?-O-glucosidase, desulfo-glucosinolate, enterococcus, glycosyl hydrolase, nitrile |
| URL Website | https://rdo.psu.ac.th/sjstweb/index.php |
| ISSN | 0125-3395 |
| Abstract | A recombinant ?-glucosidase human gut bacterium capable of nitrile production from desulfo-glucosinolates wasstudied. The bgl4 gene (2,151 bp) from Enterococcus casseliflavus CP1 was cloned and overexpressed in Escherichia coliBL21(DE3) at 25 ?C for 16 h in LB medium using 0.5 mM isopropyl ?-D-1-thiogalactopyranoside inducer. The recombinantbgl4 enzyme (79 kDa) was purified using Ni2+ affinity column chromatography. This recombinant bgl4 enzyme of the glycosylhydrolase 3 family did not degrade glucosinolates; however, it transformed desulfo-glucosinolates, except for desulfoglucoraphanin, to produce the corresponding pure nitriles in citrate phosphate buffer pH 7.0 and LB medium. The bgl4 enzymeactivity toward pNPG in buffer was optimal at pH 7.0 and 37 ?C at 23.4 U/mg, and promoted by Mn2+; however, activity wasslightly deactivated by Fe2+. This provided a possible alternative metabolic route involving nitrile formation from desulfoglucosinolates by ?-glucosidase in certain bacteria. |
