| Autor: |
Sah, Balendra, Subban, Kamalraj, Jayabaskaran, Chelliah |
| Předmět: |
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| Zdroj: |
FEMS Microbiology Letters; Apr2019, Vol. 366 Issue 7, pN.PAG-N.PAG, 1p, 1 Color Photograph, 1 Diagram, 2 Graphs |
| Abstrakt: |
10-deacetylbaccatin III-10-β- O -acetyltransferase (DBAT) is a key rate-limiting enzyme of the Taxol biosynthetic pathway, which is uncharacterized in Taxol-producing endophytic fungi. Here, an open reading frame of DBAT was cloned from the Taxol-producing endophytic fungus Lasiodiplodia theobromae (Lt DBAT). The Lt DBAT enzyme was heterologously expressed and purified by the affinity and gel filtration chromatography methods. The molecular weight of the purified protein was 49 kDa and its identity was confirmed by western blot. The purified Lt DBAT enzyme was capable of catalyzing 10-deacetylbaccatin III into baccatin III, as shown by liquid chromatography–mass spectroscopy. The mass spectra of baccatin III were identical to the authentic baccatin III. The Lt DBAT enzyme was characterized and the kinetic parameters of catalysis were determined. In addition, localization of Lt DBAT was performed by using confocal microscopy and the result showed that the enzyme was localized in lipid droplets. Together, this study provides biochemical insights into the fungal recombinant DBAT enzyme that is involved in the Taxol biosynthetic pathway. In the near future, engineering of the Lt DBAT enzyme and the Taxol biosynthetic pathway in endophytic fungi could be an eco-friendly and economically feasible alternative source for production of Taxol and its precursors. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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