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Updated: Jun 4, 2026
10:09
A Straightforward Method for Glucosinolate Extraction and Analysis with High-pressure Liquid Chromatography (HPLC)
Published on: March 15, 2017
Modulation of sulfur metabolism enables efficient glucosinolate engineering.
Morten E Møldrup1, Fernando Geu-Flores, Carl E Olsen
1Department of Plant Biology and Biotechnology, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark.
BMC Biotechnology
|February 2, 2011
Summary
Metabolic engineering in plants can produce cancer-preventive benzylglucosinolates (BGLS). Enhancing sulfur metabolism via adenosine 5'-phosphosulfate kinase 2 (APK2) significantly boosted BGLS production in Nicotiana benthamiana.
Area of Science:
- Plant biotechnology
- Metabolic engineering
- Natural product biosynthesis
Background:
- Metabolic engineering aims to produce valuable natural products in heterologous hosts.
- Glucosinolates, found in cruciferous plants, are known for their cancer-preventive properties.
- Previous work established benzylglucosinolate (BGLS) production in Nicotiana benthamiana, but accumulation was suboptimal.
Purpose of the Study:
- To optimize heterologous benzylglucosinolate (BGLS) production in Nicotiana benthamiana.
- To identify and overcome metabolic bottlenecks in the BGLS biosynthesis pathway.
- To enhance the efficiency of glucosinolate production in non-cruciferous plants.
Main Methods:
- Comparative metabolite analysis of BGLS-producing N. benthamiana leaves.
- Investigating the role of γ-glutamyl peptidase 1 (GGP1) and sulfotransferase AtSOT16.
- Evaluating the impact of genes involved in 3 -phosphoadenosine-5 -phosphosulfate (PAPS) metabolism, including adenosine 5 -phosphosulfate kinase 2 (APK2).
Main Results:
- Increased BGLS levels with GGP1 were associated with accumulation of the intermediate desulfoBGLS.
- A bottleneck was identified in the sulfation step catalyzed by AtSOT16.
- Co-expression of adenosine 5 -phosphosulfate kinase 2 (APK2) reduced desulfoBGLS accumulation by over 98% and increased BGLS by 16-fold.
Conclusions:
- Optimizing sulfur metabolism by redirecting sulfur from primary to secondary metabolism significantly improves BGLS accumulation.
- This study presents a key advancement for clean and efficient glucosinolate production in heterologous systems.
- The findings highlight the critical importance of considering co-substrates and their biological roles in metabolic engineering endeavors.
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