De novo production of benzyl glucosinolate in Escherichia coli

Annette Petersen; Christoph Crocoll; Barbara Ann Halkier

doi:10.1016/j.ymben.2019.02.004

De novo production of benzyl glucosinolate in Escherichia coli

Annette Petersen, Christoph Crocoll, Barbara Ann Halkier^*

^*Corresponding author af dette arbejde

Molekylær Plantebiologi

11 Citationer (Scopus)

22 Downloads (Pure)

Abstract

Microbial production of plant specialised metabolites is challenging as the biosynthetic pathways are often complex and can contain enzymes, which function is not supported in traditional production hosts. Glucosinolates are specialised metabolites of strong commercial interest due to their health-promoting effects. In this work, we engineered the production of benzyl glucosinolate in Escherichia coli. We systematically optimised the production levels by first screening different expression strains and by modification of growth conditions and media compositions. This resulted in production from undetectable to approximately 4.1 μM benzyl glucosinolate, but also approximately 3.7 μM of desulfo-benzyl glucosinolate, the final intermediate of this pathway. Additional optimisation of pathway flux through entry point cytochrome P450 enzymes and PAPS-dependent sulfotransferase increased the production additionally 5-fold to 20.3 μM (equivalent to 8.3 mg/L) benzyl glucosinolate.

Originalsprog	Engelsk
Tidsskrift	Metabolic Engineering
Vol/bind	54
Sider (fra-til)	24-34
ISSN	1096-7176
DOI	https://doi.org/10.1016/j.ymben.2019.02.004
Status	Udgivet - 1 jul. 2019

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10.1016/j.ymben.2019.02.004Licens: CC BY-NC-ND

De novo production of benzyl glucosinolate in Escherichia coliForlagets udgivne version, 1,3 MBLicens: CC BY-NC-ND

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title = "De novo production of benzyl glucosinolate in Escherichia coli",

abstract = "Microbial production of plant specialised metabolites is challenging as the biosynthetic pathways are often complex and can contain enzymes, which function is not supported in traditional production hosts. Glucosinolates are specialised metabolites of strong commercial interest due to their health-promoting effects. In this work, we engineered the production of benzyl glucosinolate in Escherichia coli. We systematically optimised the production levels by first screening different expression strains and by modification of growth conditions and media compositions. This resulted in production from undetectable to approximately 4.1 μM benzyl glucosinolate, but also approximately 3.7 μM of desulfo-benzyl glucosinolate, the final intermediate of this pathway. Additional optimisation of pathway flux through entry point cytochrome P450 enzymes and PAPS-dependent sulfotransferase increased the production additionally 5-fold to 20.3 μM (equivalent to 8.3 mg/L) benzyl glucosinolate.",

keywords = "Escherichia coli, Glucosinolates, Metabolic engineering, Targeted proteomics",

author = "Annette Petersen and Christoph Crocoll and Halkier, {Barbara Ann}",

year = "2019",

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doi = "10.1016/j.ymben.2019.02.004",

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journal = "Metabolic Engineering",

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TY - JOUR

T1 - De novo production of benzyl glucosinolate in Escherichia coli

AU - Petersen, Annette

AU - Crocoll, Christoph

AU - Halkier, Barbara Ann

PY - 2019/7/1

Y1 - 2019/7/1

N2 - Microbial production of plant specialised metabolites is challenging as the biosynthetic pathways are often complex and can contain enzymes, which function is not supported in traditional production hosts. Glucosinolates are specialised metabolites of strong commercial interest due to their health-promoting effects. In this work, we engineered the production of benzyl glucosinolate in Escherichia coli. We systematically optimised the production levels by first screening different expression strains and by modification of growth conditions and media compositions. This resulted in production from undetectable to approximately 4.1 μM benzyl glucosinolate, but also approximately 3.7 μM of desulfo-benzyl glucosinolate, the final intermediate of this pathway. Additional optimisation of pathway flux through entry point cytochrome P450 enzymes and PAPS-dependent sulfotransferase increased the production additionally 5-fold to 20.3 μM (equivalent to 8.3 mg/L) benzyl glucosinolate.

AB - Microbial production of plant specialised metabolites is challenging as the biosynthetic pathways are often complex and can contain enzymes, which function is not supported in traditional production hosts. Glucosinolates are specialised metabolites of strong commercial interest due to their health-promoting effects. In this work, we engineered the production of benzyl glucosinolate in Escherichia coli. We systematically optimised the production levels by first screening different expression strains and by modification of growth conditions and media compositions. This resulted in production from undetectable to approximately 4.1 μM benzyl glucosinolate, but also approximately 3.7 μM of desulfo-benzyl glucosinolate, the final intermediate of this pathway. Additional optimisation of pathway flux through entry point cytochrome P450 enzymes and PAPS-dependent sulfotransferase increased the production additionally 5-fold to 20.3 μM (equivalent to 8.3 mg/L) benzyl glucosinolate.

KW - Escherichia coli

KW - Glucosinolates

KW - Metabolic engineering

KW - Targeted proteomics

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SN - 1096-7176

VL - 54

SP - 24

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JO - Metabolic Engineering

JF - Metabolic Engineering

ER -

De novo production of benzyl glucosinolate in Escherichia coli

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