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Biochemical insights into the recombinant 10-deacetylbaccatin III-10-β-O-acetyltransferase enzyme from the Taxol-producing endophytic fungus Lasiodiplodia theobromae
by
Jayabaskaran, Chelliah
, Subban, Kamalraj
, Sah, Balendra
in
Acetyltransferase
/ Acetyltransferases - chemistry
/ Acetyltransferases - genetics
/ Acetyltransferases - metabolism
/ Alkaloids - metabolism
/ Ascomycota - chemistry
/ Ascomycota - enzymology
/ Ascomycota - genetics
/ Ascomycota - metabolism
/ Biocatalysis
/ biochemical pathways
/ Catalysis
/ catalytic activity
/ Chromatography
/ Cloning, Molecular
/ Confocal microscopy
/ droplets
/ economic feasibility
/ Endophytes
/ Endophytes - enzymology
/ Endophytes - genetics
/ Endophytes - metabolism
/ Enzyme Stability
/ Enzymes
/ Fungal Proteins - chemistry
/ Fungal Proteins - genetics
/ Fungal Proteins - metabolism
/ Fungi
/ gel chromatography
/ Gel filtration
/ Hydrogen-Ion Concentration
/ Kinetics
/ Lasiodiplodia theobromae
/ Lipids
/ Liquid chromatography
/ Localization
/ Mass spectra
/ mass spectrometry
/ Mass spectroscopy
/ Microbiology
/ Molecular weight
/ open reading frames
/ Paclitaxel
/ Paclitaxel - biosynthesis
/ Recombinant Proteins - chemistry
/ Recombinant Proteins - genetics
/ Recombinant Proteins - metabolism
/ Spectrum analysis
/ Taxoids - metabolism
/ Taxol
/ Temperature
/ transferases
/ Western blotting
/ Yeast
2019
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Biochemical insights into the recombinant 10-deacetylbaccatin III-10-β-O-acetyltransferase enzyme from the Taxol-producing endophytic fungus Lasiodiplodia theobromae
by
Jayabaskaran, Chelliah
, Subban, Kamalraj
, Sah, Balendra
in
Acetyltransferase
/ Acetyltransferases - chemistry
/ Acetyltransferases - genetics
/ Acetyltransferases - metabolism
/ Alkaloids - metabolism
/ Ascomycota - chemistry
/ Ascomycota - enzymology
/ Ascomycota - genetics
/ Ascomycota - metabolism
/ Biocatalysis
/ biochemical pathways
/ Catalysis
/ catalytic activity
/ Chromatography
/ Cloning, Molecular
/ Confocal microscopy
/ droplets
/ economic feasibility
/ Endophytes
/ Endophytes - enzymology
/ Endophytes - genetics
/ Endophytes - metabolism
/ Enzyme Stability
/ Enzymes
/ Fungal Proteins - chemistry
/ Fungal Proteins - genetics
/ Fungal Proteins - metabolism
/ Fungi
/ gel chromatography
/ Gel filtration
/ Hydrogen-Ion Concentration
/ Kinetics
/ Lasiodiplodia theobromae
/ Lipids
/ Liquid chromatography
/ Localization
/ Mass spectra
/ mass spectrometry
/ Mass spectroscopy
/ Microbiology
/ Molecular weight
/ open reading frames
/ Paclitaxel
/ Paclitaxel - biosynthesis
/ Recombinant Proteins - chemistry
/ Recombinant Proteins - genetics
/ Recombinant Proteins - metabolism
/ Spectrum analysis
/ Taxoids - metabolism
/ Taxol
/ Temperature
/ transferases
/ Western blotting
/ Yeast
2019
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Biochemical insights into the recombinant 10-deacetylbaccatin III-10-β-O-acetyltransferase enzyme from the Taxol-producing endophytic fungus Lasiodiplodia theobromae
by
Jayabaskaran, Chelliah
, Subban, Kamalraj
, Sah, Balendra
in
Acetyltransferase
/ Acetyltransferases - chemistry
/ Acetyltransferases - genetics
/ Acetyltransferases - metabolism
/ Alkaloids - metabolism
/ Ascomycota - chemistry
/ Ascomycota - enzymology
/ Ascomycota - genetics
/ Ascomycota - metabolism
/ Biocatalysis
/ biochemical pathways
/ Catalysis
/ catalytic activity
/ Chromatography
/ Cloning, Molecular
/ Confocal microscopy
/ droplets
/ economic feasibility
/ Endophytes
/ Endophytes - enzymology
/ Endophytes - genetics
/ Endophytes - metabolism
/ Enzyme Stability
/ Enzymes
/ Fungal Proteins - chemistry
/ Fungal Proteins - genetics
/ Fungal Proteins - metabolism
/ Fungi
/ gel chromatography
/ Gel filtration
/ Hydrogen-Ion Concentration
/ Kinetics
/ Lasiodiplodia theobromae
/ Lipids
/ Liquid chromatography
/ Localization
/ Mass spectra
/ mass spectrometry
/ Mass spectroscopy
/ Microbiology
/ Molecular weight
/ open reading frames
/ Paclitaxel
/ Paclitaxel - biosynthesis
/ Recombinant Proteins - chemistry
/ Recombinant Proteins - genetics
/ Recombinant Proteins - metabolism
/ Spectrum analysis
/ Taxoids - metabolism
/ Taxol
/ Temperature
/ transferases
/ Western blotting
/ Yeast
2019
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Biochemical insights into the recombinant 10-deacetylbaccatin III-10-β-O-acetyltransferase enzyme from the Taxol-producing endophytic fungus Lasiodiplodia theobromae
Journal Article
Biochemical insights into the recombinant 10-deacetylbaccatin III-10-β-O-acetyltransferase enzyme from the Taxol-producing endophytic fungus Lasiodiplodia theobromae
2019
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Overview
ABSTRACT
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 (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.
Biochemical insights into the biotechnologically important fungal recombinant 10-deacetylbaccatin III-10-I2-O-acetyltransferase enzyme.
Publisher
Oxford University Press
Subject
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