De novo production of benzyl glucosinolate in Escherichia coli

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

UR - http://www.scopus.com/inward/record.url?scp=85063026606&partnerID=8YFLogxK

U2 - 10.1016/j.ymben.2019.02.004

DO - 10.1016/j.ymben.2019.02.004

M3 - Journal article

C2 - 30831267

AN - SCOPUS:85063026606

SN - 1096-7176

VL - 54

SP - 24

EP - 34

JO - Metabolic Engineering

JF - Metabolic Engineering

ER -