An improved Nicotiana benthamiana bioproduction chassis provides novel insights into nicotine biosynthesis

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

Standard

An improved Nicotiana benthamiana bioproduction chassis provides novel insights into nicotine biosynthesis. / Vollheyde, Katharina; Dudley, Quentin M.; Yang, Ting; Oz, Mehmet T.; Mancinotti, Davide; Fedi, Mariano Olivera; Heavens, Darren; Linsmith, Gareth; Chhetry, Monika; Smedley, Mark A.; Harwood, Wendy A.; Swarbreck, David; Geu-Flores, Fernando; Patron, Nicola J.

In: New Phytologist, Vol. 240, No. 1, 2023, p. 302-317.

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

Harvard

Vollheyde, K, Dudley, QM, Yang, T, Oz, MT, Mancinotti, D, Fedi, MO, Heavens, D, Linsmith, G, Chhetry, M, Smedley, MA, Harwood, WA, Swarbreck, D, Geu-Flores, F & Patron, NJ 2023, 'An improved Nicotiana benthamiana bioproduction chassis provides novel insights into nicotine biosynthesis', New Phytologist, vol. 240, no. 1, pp. 302-317. https://doi.org/10.1111/nph.19141

APA

Vollheyde, K., Dudley, Q. M., Yang, T., Oz, M. T., Mancinotti, D., Fedi, M. O., Heavens, D., Linsmith, G., Chhetry, M., Smedley, M. A., Harwood, W. A., Swarbreck, D., Geu-Flores, F., & Patron, N. J. (2023). An improved Nicotiana benthamiana bioproduction chassis provides novel insights into nicotine biosynthesis. New Phytologist, 240(1), 302-317. https://doi.org/10.1111/nph.19141

Vancouver

Vollheyde K, Dudley QM, Yang T, Oz MT, Mancinotti D, Fedi MO et al. An improved Nicotiana benthamiana bioproduction chassis provides novel insights into nicotine biosynthesis. New Phytologist. 2023;240(1):302-317. https://doi.org/10.1111/nph.19141

Author

Vollheyde, Katharina ; Dudley, Quentin M. ; Yang, Ting ; Oz, Mehmet T. ; Mancinotti, Davide ; Fedi, Mariano Olivera ; Heavens, Darren ; Linsmith, Gareth ; Chhetry, Monika ; Smedley, Mark A. ; Harwood, Wendy A. ; Swarbreck, David ; Geu-Flores, Fernando ; Patron, Nicola J. / An improved Nicotiana benthamiana bioproduction chassis provides novel insights into nicotine biosynthesis. In: New Phytologist. 2023 ; Vol. 240, No. 1. pp. 302-317.

Bibtex

@article{615d9806bf59400a81e8fd8a0b34701d,
title = "An improved Nicotiana benthamiana bioproduction chassis provides novel insights into nicotine biosynthesis",
abstract = "The model plant Nicotiana benthamiana is an increasingly attractive organism for the production of high-value, biologically active molecules. However, N. benthamiana accumulates high levels of pyridine alkaloids, in particular nicotine, which complicates the downstream purification processes. Here, we report a new assembly of the N. benthamiana genome as well as the generation of low-nicotine lines by CRISPR/Cas9-based inactivation of berberine bridge enzyme-like proteins (BBLs). Triple as well as quintuple mutants accumulated three to four times less nicotine than the respective control lines. The availability of lines without functional BBLs allowed us to probe their catalytic role in nicotine biosynthesis, which has remained obscure. Notably, chiral analysis revealed that the enantiomeric purity of nicotine was fully lost in the quintuple mutants. In addition, precursor feeding experiments showed that these mutants cannot facilitate the specific loss of C6 hydrogen that characterizes natural nicotine biosynthesis. Our work delivers an improved N. benthamiana chassis for bioproduction and uncovers the crucial role of BBLs in the stereoselectivity of nicotine biosynthesis.",
keywords = "alkaloid biosynthesis, gene editing, metabolic engineering, Nicotiana benthamiana, nicotine",
author = "Katharina Vollheyde and Dudley, {Quentin M.} and Ting Yang and Oz, {Mehmet T.} and Davide Mancinotti and Fedi, {Mariano Olivera} and Darren Heavens and Gareth Linsmith and Monika Chhetry and Smedley, {Mark A.} and Harwood, {Wendy A.} and David Swarbreck and Fernando Geu-Flores and Patron, {Nicola J.}",
note = "Publisher Copyright: {\textcopyright} 2023 The Authors. New Phytologist {\textcopyright} 2023 New Phytologist Foundation.",
year = "2023",
doi = "10.1111/nph.19141",
language = "English",
volume = "240",
pages = "302--317",
journal = "New Phytologist",
issn = "0028-646X",
publisher = "Academic Press",
number = "1",

}

RIS

TY - JOUR

T1 - An improved Nicotiana benthamiana bioproduction chassis provides novel insights into nicotine biosynthesis

AU - Vollheyde, Katharina

AU - Dudley, Quentin M.

AU - Yang, Ting

AU - Oz, Mehmet T.

AU - Mancinotti, Davide

AU - Fedi, Mariano Olivera

AU - Heavens, Darren

AU - Linsmith, Gareth

AU - Chhetry, Monika

AU - Smedley, Mark A.

AU - Harwood, Wendy A.

AU - Swarbreck, David

AU - Geu-Flores, Fernando

AU - Patron, Nicola J.

N1 - Publisher Copyright: © 2023 The Authors. New Phytologist © 2023 New Phytologist Foundation.

PY - 2023

Y1 - 2023

N2 - The model plant Nicotiana benthamiana is an increasingly attractive organism for the production of high-value, biologically active molecules. However, N. benthamiana accumulates high levels of pyridine alkaloids, in particular nicotine, which complicates the downstream purification processes. Here, we report a new assembly of the N. benthamiana genome as well as the generation of low-nicotine lines by CRISPR/Cas9-based inactivation of berberine bridge enzyme-like proteins (BBLs). Triple as well as quintuple mutants accumulated three to four times less nicotine than the respective control lines. The availability of lines without functional BBLs allowed us to probe their catalytic role in nicotine biosynthesis, which has remained obscure. Notably, chiral analysis revealed that the enantiomeric purity of nicotine was fully lost in the quintuple mutants. In addition, precursor feeding experiments showed that these mutants cannot facilitate the specific loss of C6 hydrogen that characterizes natural nicotine biosynthesis. Our work delivers an improved N. benthamiana chassis for bioproduction and uncovers the crucial role of BBLs in the stereoselectivity of nicotine biosynthesis.

AB - The model plant Nicotiana benthamiana is an increasingly attractive organism for the production of high-value, biologically active molecules. However, N. benthamiana accumulates high levels of pyridine alkaloids, in particular nicotine, which complicates the downstream purification processes. Here, we report a new assembly of the N. benthamiana genome as well as the generation of low-nicotine lines by CRISPR/Cas9-based inactivation of berberine bridge enzyme-like proteins (BBLs). Triple as well as quintuple mutants accumulated three to four times less nicotine than the respective control lines. The availability of lines without functional BBLs allowed us to probe their catalytic role in nicotine biosynthesis, which has remained obscure. Notably, chiral analysis revealed that the enantiomeric purity of nicotine was fully lost in the quintuple mutants. In addition, precursor feeding experiments showed that these mutants cannot facilitate the specific loss of C6 hydrogen that characterizes natural nicotine biosynthesis. Our work delivers an improved N. benthamiana chassis for bioproduction and uncovers the crucial role of BBLs in the stereoselectivity of nicotine biosynthesis.

KW - alkaloid biosynthesis

KW - gene editing

KW - metabolic engineering

KW - Nicotiana benthamiana

KW - nicotine

U2 - 10.1111/nph.19141

DO - 10.1111/nph.19141

M3 - Journal article

C2 - 37488711

AN - SCOPUS:85165581637

VL - 240

SP - 302

EP - 317

JO - New Phytologist

JF - New Phytologist

SN - 0028-646X

IS - 1

ER -

ID: 363060517