Publication

Metabolic engineering of bacillus subtilis toward taxadiene biosynthesis as the first committed step for taxol production

Abdallah, I. I., Pramastya, H., Van Merkerk, R., Sukrasno & Quax, W. J., 1-Jun-2019, In : Frontiers in Microbiology. 10, FEB, 11 p., 218.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Abdallah, I. I., Pramastya, H., Van Merkerk, R., Sukrasno, & Quax, W. J. (2019). Metabolic engineering of bacillus subtilis toward taxadiene biosynthesis as the first committed step for taxol production. Frontiers in Microbiology, 10(FEB), [218]. https://doi.org/10.3389/fmicb.2019.00218

Author

Abdallah, Ingy I. ; Pramastya, Hegar ; Van Merkerk, Ronald ; Sukrasno, ; Quax, Wim J. / Metabolic engineering of bacillus subtilis toward taxadiene biosynthesis as the first committed step for taxol production. In: Frontiers in Microbiology. 2019 ; Vol. 10, No. FEB.

Harvard

Abdallah, II, Pramastya, H, Van Merkerk, R, Sukrasno, & Quax, WJ 2019, 'Metabolic engineering of bacillus subtilis toward taxadiene biosynthesis as the first committed step for taxol production', Frontiers in Microbiology, vol. 10, no. FEB, 218. https://doi.org/10.3389/fmicb.2019.00218

Standard

Metabolic engineering of bacillus subtilis toward taxadiene biosynthesis as the first committed step for taxol production. / Abdallah, Ingy I.; Pramastya, Hegar; Van Merkerk, Ronald; Sukrasno, ; Quax, Wim J.

In: Frontiers in Microbiology, Vol. 10, No. FEB, 218, 01.06.2019.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Abdallah II, Pramastya H, Van Merkerk R, Sukrasno , Quax WJ. Metabolic engineering of bacillus subtilis toward taxadiene biosynthesis as the first committed step for taxol production. Frontiers in Microbiology. 2019 Jun 1;10(FEB). 218. https://doi.org/10.3389/fmicb.2019.00218


BibTeX

@article{77fc078bd2074163847edffe66ad95ec,
title = "Metabolic engineering of bacillus subtilis toward taxadiene biosynthesis as the first committed step for taxol production",
abstract = "Terpenoids are natural products known for their medicinal and commercial applications. Metabolic engineering of microbial hosts for the production of valuable compounds, such as artemisinin and Taxol, has gained vast interest in the last few decades. The Generally Regarded As Safe (GRAS) Bacillus subtilis 168 with its broad metabolic potential is considered one of these interesting microbial hosts. In the effort toward engineering B. subtilis as a cell factory for the production of the chemotherapeutic Taxol, we expressed the plant-derived taxadiene synthase (TXS) enzyme. TXS is responsible for the conversion of the precursor geranylgeranyl pyrophosphate (GGPP) to taxa-4,11-diene, which is the first committed intermediate in Taxol biosynthesis. Furthermore, overexpression of eight enzymes in the biosynthesis pathway was performed to increase the flux of the GGPP precursor. This was achieved by creating a synthetic operon harboring the B. subtilis genes encoding the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway (dxs, ispD, ispF, ispH, ispC, ispE, ispG) together with ispA (encoding geranyl and farnesyl pyrophosphate synthases) responsible for providing farnesyl pyrophosphate (FPP). In addition, a vector harboring the crtE gene (encoding geranylgeranyl pyrophosphate synthase, GGPPS, of Pantoea ananatis) to increase the supply of GGPP was introduced. The overexpression of the MEP pathway enzymes along with IspA and GGPPS caused an 83-fold increase in the amount of taxadiene produced compared to the strain only expressing TXS and relying on the innate pathway of B. subtilis. The total amount of taxadiene produced by that strain was 17.8 mg/l. This is the first account of the successful expression of taxadiene synthase in B. subtilis. We determined that the expression of GGPPS through the crtE gene is essential for the formation of sufficient precursor, GGPP, in B. subtilis as its innate metabolism is not efficient in producing it. Finally, the extracellular localization of taxadiene production by overexpressing the complete MEP pathway along with IspA and GGPPS presents the prospect for further engineering aiming for semisynthesis of Taxol.",
keywords = "Bacillus subtilis, metabolite, MEP, GGPPS, taxadiene, Taxol, PYROPHOSPHATE SYNTHETASE, ESCHERICHIA-COLI, EXPRESSION, PACLITAXEL, PRECURSOR, SYNTHASE, PATHWAY, GENES, INTERMEDIATE, OPTIMIZATION",
author = "Abdallah, {Ingy I.} and Hegar Pramastya and {Van Merkerk}, Ronald and Sukrasno and Quax, {Wim J.}",
year = "2019",
month = "6",
day = "1",
doi = "10.3389/fmicb.2019.00218",
language = "English",
volume = "10",
journal = "Frontiers in Microbiology",
issn = "1664-302X",
publisher = "Frontiers Media S.A.",
number = "FEB",

}

RIS

TY - JOUR

T1 - Metabolic engineering of bacillus subtilis toward taxadiene biosynthesis as the first committed step for taxol production

AU - Abdallah, Ingy I.

AU - Pramastya, Hegar

AU - Van Merkerk, Ronald

AU - Sukrasno, null

AU - Quax, Wim J.

PY - 2019/6/1

Y1 - 2019/6/1

N2 - Terpenoids are natural products known for their medicinal and commercial applications. Metabolic engineering of microbial hosts for the production of valuable compounds, such as artemisinin and Taxol, has gained vast interest in the last few decades. The Generally Regarded As Safe (GRAS) Bacillus subtilis 168 with its broad metabolic potential is considered one of these interesting microbial hosts. In the effort toward engineering B. subtilis as a cell factory for the production of the chemotherapeutic Taxol, we expressed the plant-derived taxadiene synthase (TXS) enzyme. TXS is responsible for the conversion of the precursor geranylgeranyl pyrophosphate (GGPP) to taxa-4,11-diene, which is the first committed intermediate in Taxol biosynthesis. Furthermore, overexpression of eight enzymes in the biosynthesis pathway was performed to increase the flux of the GGPP precursor. This was achieved by creating a synthetic operon harboring the B. subtilis genes encoding the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway (dxs, ispD, ispF, ispH, ispC, ispE, ispG) together with ispA (encoding geranyl and farnesyl pyrophosphate synthases) responsible for providing farnesyl pyrophosphate (FPP). In addition, a vector harboring the crtE gene (encoding geranylgeranyl pyrophosphate synthase, GGPPS, of Pantoea ananatis) to increase the supply of GGPP was introduced. The overexpression of the MEP pathway enzymes along with IspA and GGPPS caused an 83-fold increase in the amount of taxadiene produced compared to the strain only expressing TXS and relying on the innate pathway of B. subtilis. The total amount of taxadiene produced by that strain was 17.8 mg/l. This is the first account of the successful expression of taxadiene synthase in B. subtilis. We determined that the expression of GGPPS through the crtE gene is essential for the formation of sufficient precursor, GGPP, in B. subtilis as its innate metabolism is not efficient in producing it. Finally, the extracellular localization of taxadiene production by overexpressing the complete MEP pathway along with IspA and GGPPS presents the prospect for further engineering aiming for semisynthesis of Taxol.

AB - Terpenoids are natural products known for their medicinal and commercial applications. Metabolic engineering of microbial hosts for the production of valuable compounds, such as artemisinin and Taxol, has gained vast interest in the last few decades. The Generally Regarded As Safe (GRAS) Bacillus subtilis 168 with its broad metabolic potential is considered one of these interesting microbial hosts. In the effort toward engineering B. subtilis as a cell factory for the production of the chemotherapeutic Taxol, we expressed the plant-derived taxadiene synthase (TXS) enzyme. TXS is responsible for the conversion of the precursor geranylgeranyl pyrophosphate (GGPP) to taxa-4,11-diene, which is the first committed intermediate in Taxol biosynthesis. Furthermore, overexpression of eight enzymes in the biosynthesis pathway was performed to increase the flux of the GGPP precursor. This was achieved by creating a synthetic operon harboring the B. subtilis genes encoding the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway (dxs, ispD, ispF, ispH, ispC, ispE, ispG) together with ispA (encoding geranyl and farnesyl pyrophosphate synthases) responsible for providing farnesyl pyrophosphate (FPP). In addition, a vector harboring the crtE gene (encoding geranylgeranyl pyrophosphate synthase, GGPPS, of Pantoea ananatis) to increase the supply of GGPP was introduced. The overexpression of the MEP pathway enzymes along with IspA and GGPPS caused an 83-fold increase in the amount of taxadiene produced compared to the strain only expressing TXS and relying on the innate pathway of B. subtilis. The total amount of taxadiene produced by that strain was 17.8 mg/l. This is the first account of the successful expression of taxadiene synthase in B. subtilis. We determined that the expression of GGPPS through the crtE gene is essential for the formation of sufficient precursor, GGPP, in B. subtilis as its innate metabolism is not efficient in producing it. Finally, the extracellular localization of taxadiene production by overexpressing the complete MEP pathway along with IspA and GGPPS presents the prospect for further engineering aiming for semisynthesis of Taxol.

KW - Bacillus subtilis

KW - metabolite

KW - MEP

KW - GGPPS

KW - taxadiene

KW - Taxol

KW - PYROPHOSPHATE SYNTHETASE

KW - ESCHERICHIA-COLI

KW - EXPRESSION

KW - PACLITAXEL

KW - PRECURSOR

KW - SYNTHASE

KW - PATHWAY

KW - GENES

KW - INTERMEDIATE

KW - OPTIMIZATION

U2 - 10.3389/fmicb.2019.00218

DO - 10.3389/fmicb.2019.00218

M3 - Article

VL - 10

JO - Frontiers in Microbiology

JF - Frontiers in Microbiology

SN - 1664-302X

IS - FEB

M1 - 218

ER -

ID: 76907162