Integrating pathway elucidation with yeast engineering to produce polpunonic acid the precursor of the anti-obesity agent celastrol

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Integrating pathway elucidation with yeast engineering to produce polpunonic acid the precursor of the anti-obesity agent celastrol. / Hansen, Nikolaj Lervad; Miettinen, Karel; Zhao, Yong; Ignea, Codruta; Andreadeli, Aggeliki; Raadam, Morten Hessellund; Makris, Antonios M.; Møller, Birger Lindberg; Stærk, Dan; Bak, Søren; Kampranis, Sotirios.

In: Microbial Cell Factories, Vol. 19, 15, 2020.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Hansen, NL, Miettinen, K, Zhao, Y, Ignea, C, Andreadeli, A, Raadam, MH, Makris, AM, Møller, BL, Stærk, D, Bak, S & Kampranis, S 2020, 'Integrating pathway elucidation with yeast engineering to produce polpunonic acid the precursor of the anti-obesity agent celastrol', Microbial Cell Factories, vol. 19, 15. https://doi.org/10.1186/s12934-020-1284-9

APA

Hansen, N. L., Miettinen, K., Zhao, Y., Ignea, C., Andreadeli, A., Raadam, M. H., Makris, A. M., Møller, B. L., Stærk, D., Bak, S., & Kampranis, S. (2020). Integrating pathway elucidation with yeast engineering to produce polpunonic acid the precursor of the anti-obesity agent celastrol. Microbial Cell Factories, 19, [15]. https://doi.org/10.1186/s12934-020-1284-9

Vancouver

Hansen NL, Miettinen K, Zhao Y, Ignea C, Andreadeli A, Raadam MH et al. Integrating pathway elucidation with yeast engineering to produce polpunonic acid the precursor of the anti-obesity agent celastrol. Microbial Cell Factories. 2020;19. 15. https://doi.org/10.1186/s12934-020-1284-9

Author

Hansen, Nikolaj Lervad ; Miettinen, Karel ; Zhao, Yong ; Ignea, Codruta ; Andreadeli, Aggeliki ; Raadam, Morten Hessellund ; Makris, Antonios M. ; Møller, Birger Lindberg ; Stærk, Dan ; Bak, Søren ; Kampranis, Sotirios. / Integrating pathway elucidation with yeast engineering to produce polpunonic acid the precursor of the anti-obesity agent celastrol. In: Microbial Cell Factories. 2020 ; Vol. 19.

Bibtex

@article{d0a0bffe12284b71af63d2432431dfd4,
title = "Integrating pathway elucidation with yeast engineering to produce polpunonic acid the precursor of the anti-obesity agent celastrol",
abstract = "Background: Celastrol is a promising anti-obesity agent that acts as a sensitizer of the protein hormone leptin. Despite its potent activity, a sustainable source of celastrol and celastrol derivatives for further pharmacological studies is lacking.Results: To elucidate the celastrol biosynthetic pathway and reconstruct it in Saccharomyces cerevisiae, we mined a root-transcriptome of Tripterygium wilfordii and identified four oxidosqualene cyclases and 49 cytochrome P450s as candidates to be involved in the early steps of celastrol biosynthesis. Using functional screening of the candidate genes in Nicotiana benthamiana, TwOSC4 was characterized as a novel oxidosqualene cyclase that produces friedelin, the presumed triterpenoid backbone of celastrol. In addition, three P450s (CYP712K1, CYP712K2, and CYP712K3) that act downstream of TwOSC4 were found to effectively oxidize friedelin and form the likely celastrol biosynthesis intermediates 29-hydroxy-friedelin and polpunonic acid. To facilitate production of friedelin, the yeast strain AM254 was constructed by deleting UBC7, which afforded a 5-fold increase in friedelin titer. This platform was further expanded with CYP712K1 to produce polpunonic acid and a method for the facile extraction of products from the yeast culture medium, resulting in polpunonic acid titers of 1.4 mg/L.Conclusion: Our study elucidates the early steps of celastrol biosynthesis and paves the way for future biotechnological production of this pharmacologically promising compound in engineered yeast strains.",
author = "Hansen, {Nikolaj Lervad} and Karel Miettinen and Yong Zhao and Codruta Ignea and Aggeliki Andreadeli and Raadam, {Morten Hessellund} and Makris, {Antonios M.} and M{\o}ller, {Birger Lindberg} and Dan St{\ae}rk and S{\o}ren Bak and Sotirios Kampranis",
year = "2020",
doi = "10.1186/s12934-020-1284-9",
language = "English",
volume = "19",
journal = "Microbial Cell",
issn = "1475-2859",
publisher = "BioMed Central",

}

RIS

TY - JOUR

T1 - Integrating pathway elucidation with yeast engineering to produce polpunonic acid the precursor of the anti-obesity agent celastrol

AU - Hansen, Nikolaj Lervad

AU - Miettinen, Karel

AU - Zhao, Yong

AU - Ignea, Codruta

AU - Andreadeli, Aggeliki

AU - Raadam, Morten Hessellund

AU - Makris, Antonios M.

AU - Møller, Birger Lindberg

AU - Stærk, Dan

AU - Bak, Søren

AU - Kampranis, Sotirios

PY - 2020

Y1 - 2020

N2 - Background: Celastrol is a promising anti-obesity agent that acts as a sensitizer of the protein hormone leptin. Despite its potent activity, a sustainable source of celastrol and celastrol derivatives for further pharmacological studies is lacking.Results: To elucidate the celastrol biosynthetic pathway and reconstruct it in Saccharomyces cerevisiae, we mined a root-transcriptome of Tripterygium wilfordii and identified four oxidosqualene cyclases and 49 cytochrome P450s as candidates to be involved in the early steps of celastrol biosynthesis. Using functional screening of the candidate genes in Nicotiana benthamiana, TwOSC4 was characterized as a novel oxidosqualene cyclase that produces friedelin, the presumed triterpenoid backbone of celastrol. In addition, three P450s (CYP712K1, CYP712K2, and CYP712K3) that act downstream of TwOSC4 were found to effectively oxidize friedelin and form the likely celastrol biosynthesis intermediates 29-hydroxy-friedelin and polpunonic acid. To facilitate production of friedelin, the yeast strain AM254 was constructed by deleting UBC7, which afforded a 5-fold increase in friedelin titer. This platform was further expanded with CYP712K1 to produce polpunonic acid and a method for the facile extraction of products from the yeast culture medium, resulting in polpunonic acid titers of 1.4 mg/L.Conclusion: Our study elucidates the early steps of celastrol biosynthesis and paves the way for future biotechnological production of this pharmacologically promising compound in engineered yeast strains.

AB - Background: Celastrol is a promising anti-obesity agent that acts as a sensitizer of the protein hormone leptin. Despite its potent activity, a sustainable source of celastrol and celastrol derivatives for further pharmacological studies is lacking.Results: To elucidate the celastrol biosynthetic pathway and reconstruct it in Saccharomyces cerevisiae, we mined a root-transcriptome of Tripterygium wilfordii and identified four oxidosqualene cyclases and 49 cytochrome P450s as candidates to be involved in the early steps of celastrol biosynthesis. Using functional screening of the candidate genes in Nicotiana benthamiana, TwOSC4 was characterized as a novel oxidosqualene cyclase that produces friedelin, the presumed triterpenoid backbone of celastrol. In addition, three P450s (CYP712K1, CYP712K2, and CYP712K3) that act downstream of TwOSC4 were found to effectively oxidize friedelin and form the likely celastrol biosynthesis intermediates 29-hydroxy-friedelin and polpunonic acid. To facilitate production of friedelin, the yeast strain AM254 was constructed by deleting UBC7, which afforded a 5-fold increase in friedelin titer. This platform was further expanded with CYP712K1 to produce polpunonic acid and a method for the facile extraction of products from the yeast culture medium, resulting in polpunonic acid titers of 1.4 mg/L.Conclusion: Our study elucidates the early steps of celastrol biosynthesis and paves the way for future biotechnological production of this pharmacologically promising compound in engineered yeast strains.

U2 - 10.1186/s12934-020-1284-9

DO - 10.1186/s12934-020-1284-9

M3 - Journal article

C2 - 31992268

VL - 19

JO - Microbial Cell

JF - Microbial Cell

SN - 1475-2859

M1 - 15

ER -

ID: 234283597