Precursor-Directed Combinatorial Biosynthesis of Cinnamoyl, Dihydrocinnamoyl, and Benzoyl Anthranilates in Saccharomyces cerevisiae

• Published in: PloS one Vol. 10; no. 10; p. e0138972
• Main Authors: Eudes, Aymerick, Teixeira Benites, Veronica, Wang, George, Baidoo, Edward E K, Lee, Taek Soon, Keasling, Jay D, Loqué, Dominique
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 02.10.2015; Public Library of Science (PLoS)
• Subjects: Acids; Arabidopsis thaliana; Baking yeast; BASIC BIOLOGICAL SCIENCES; Benzoates; Benzoic acid; Biodiversity; Biological activity; Biological effects; Biological products; Biosynthesis; Chemical properties; Chemical synthesis; Combinatorial analysis; Combinatorial Chemistry Techniques; Dianthus caryophyllus; Identification methods; Laboratories; Ligases; Metabolic pathways; Metabolism; Metabolites; Natural products; ortho-Aminobenzoates - metabolism; Physiological aspects; Populus balsamifera; Powell, Jerome; Rhodopseudomonas palustris; Saccharomyces cerevisiae; Saccharomyces cerevisiae - metabolism; Spectrometry, Mass, Electrospray Ionization; Substrates; United States > US; San Francisco California; California
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0138972

Biological synthesis of pharmaceuticals and biochemicals offers an environmentally friendly alternative to conventional chemical synthesis. These alternative methods require the design of metabolic pathways and the identification of enzymes exhibiting adequate activities. Cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates are natural metabolites which possess beneficial activities for human health, and the search is expanding for novel derivatives that might have enhanced biological activity. For example, biosynthesis in Dianthus caryophyllus is catalyzed by hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/ benzoyltransferase (HCBT), which couples hydroxycinnamoyl-CoAs and benzoyl-CoAs to anthranilate. We recently demonstrated the potential of using yeast (Saccharomyces cerevisiae) for the biological production of a few cinnamoyl anthranilates by heterologous co-expression of 4-coumaroyl:CoA ligase from Arabidopsis thaliana (4CL5) and HCBT. Here we report that, by exploiting the substrate flexibility of both 4CL5 and HCBT, we achieved rapid biosynthesis of more than 160 cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates in yeast upon feeding with both natural and non-natural cinnamates, dihydrocinnamates, benzoates, and anthranilates. Our results demonstrate the use of enzyme promiscuity in biological synthesis to achieve high chemical diversity within a defined class of molecules. This work also points to the potential for the combinatorial biosynthesis of diverse and valuable cinnamoylated, dihydrocinnamoylated, and benzoylated products by using the versatile biological enzyme 4CL5 along with characterized cinnamoyl-CoA- and benzoyl-CoA-utilizing transferases.