Biochemical insights into the recombinant 10-deacetylbaccatin III-10-beta-O-acetyltransferase enzyme from the Taxol-producing endophytic fungus Lasiodiplodia theobromae

Sah, Balendra and Subban, Kamalraj and Jayabaskaran, Chelliah (2019) Biochemical insights into the recombinant 10-deacetylbaccatin III-10-beta-O-acetyltransferase enzyme from the Taxol-producing endophytic fungus Lasiodiplodia theobromae. In: FEMS MICROBIOLOGY LETTERS, 366 (7).

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Abstract

10-deacetylbaccatin III-10-beta-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 (LtDBAT). The LtDBAT 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 LtDBAT 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 LtDBAT enzyme was characterized and the kinetic parameters of catalysis were determined. In addition, localization of LtDBAT 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 LtDBAT 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.

Item Type:	Journal Article
Publication:	FEMS MICROBIOLOGY LETTERS
Publisher:	OXFORD UNIV PRESS
Additional Information:	copyright of this article belongs to OXFORD UNIV PRESS
Keywords:	DBAT; baccatin III; Taxol; Lasiodiplodia theobromae; endophytic fungi
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	26 Jun 2019 17:23
Last Modified:	26 Jun 2019 17:23
URI:	http://eprints.iisc.ac.in/id/eprint/63084

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