Microbial production of indolylglucosinolate through engineering of a multi-gene pathway in a versatile yeast expression platform.
Epidemiological studies have shown that consumption of cruciferous vegetables, such as, broccoli and cabbages, is associated with a reduced risk of developing cancer. This phenomenon has been attributed to specific glucosinolates among the ~30 glucosinolates that are typically present as natural products characteristic of cruciferous plants. Accordingly, there has been a strong interest to produce these compounds in microbial cell factories as it will allow production of selected beneficial glucosinolates. We have developed a versatile platform for stable expression of multi-gene pathways in the yeast, Saccharomyces cerevisiae. Introduction of the seven-step pathway of indolylglucosinolate from Arabidopsis thaliana to yeast resulted in the first successful production of glucosinolates in a microbial host. The production of indolylglucosinolate was further optimized by substituting supporting endogenous yeast activities with plant-derived enzymes. Production of indolylglucosinolate serves as a proof-of-concept for our expression platform, and provides a basis for large-scale microbial production of specific glucosinolates for the benefit of human health.
University of Copenhagen, Faculty of Life Sciences, Department of Plant Biology and Biotechnology, Molecular Plant Biology, VKR Research Centre for Pro-Active Plants, 40 Thorvaldsensvej, Frederiksberg C, Copenhagen, Denmark.
Michael Dalgaard
MD
Mikkelsen
Line Due
LD
Buron
Bo
B
Salomonsen
Carl Erik
CE
Olsen
Bjarne Gram
BG
Hansen
Uffe Hasbro
UH
Mortensen
Barbara Ann
BA
Halkier
D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000096:biosynthesis* / Q000235:genetics; D001937:Brassica / Q000737:chemistry; D005961:Glucosinolates / Q000096:biosynthesis* / Q000737:chemistry; D006801:Humans; D060847:Metabolic Engineering; D009369:Neoplasms / Q000453:epidemiology / Q000517:prevention & control; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics
D029681:Arabidopsis Proteins; D005961:Glucosinolates; D011994:Recombinant Proteins