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Published on: October 4, 2019
Engineering an Escherichia coli - Saccharomyces cerevisiae consortium for 2-phenylethyl glucosinolate biosynthesis
William Thomas Wajn1, Michal Poborsky1, Christoph Crocoll1
1DynaMo Center, Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg, Denmark.
Journal of Experimental Botany
|June 2, 2026
Summary
This study presents the first microbial biosynthesis of 2-phenylethyl glucosinolate (2PE) using engineered E. coli and S. cerevisiae. This breakthrough enables the production of complex glucosinolates from amino acids, advancing nutraceutical and agricultural biotechnology.
Area of Science:
- Biotechnology
- Metabolic Engineering
- Synthetic Biology
Background:
- Glucosinolates are vital sulfur-rich plant metabolites with significant nutraceutical and agricultural applications.
- Microbial production of complex glucosinolates is hindered by challenges in expressing key biosynthetic enzymes, particularly those involved in chain elongation of amino acids.
Purpose of the Study:
- To establish the first microbial biosynthesis pathway for 2-phenylethyl glucosinolate (2PE), a complex glucosinolate derived from chain-elongated amino acids.
- To develop a modular engineering strategy combining Escherichia coli and Saccharomyces cerevisiae for efficient production.
Main Methods:
- Utilized a modular engineering approach, with a bacterial module for amino acid side chain elongation (phenylalanine to homophenylalanine) and a yeast module for core glucosinolate structure synthesis.
- Optimized individual modules by screening brassicaceous enzyme variants (CYP79F, MAM, BCAT) and enhanced co-factor availability (PAPS) through sulfate feeding.
- Implemented co-culture strategies, optimized carbon sources, and autoinduction for enhanced 2PE titers.
Main Results:
- Successfully achieved functional expression of an iron-sulfur cluster enzyme in E. coli for amino acid elongation.
- Identified superior enzyme variants and optimized PAPS co-factor availability, leading to a 10-fold increase in 2PE titers and alleviating the desulfo-2-phenylethyl glucosinolate (ds-2PE) bottleneck.
- Demonstrated the first microbial biosynthesis of 2PE from phenylalanine by combining the engineered bacterial and yeast modules.
Conclusions:
- Established a robust platform for the microbial production of complex glucosinolates derived from chain-elongated amino acids.
- Pathway modularization and co-culture engineering are effective strategies for overcoming limitations in expressing complex biosynthetic pathways.
- This work paves the way for biotechnological applications of glucosinolates in nutraceuticals and agriculture.
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