title: Precursor-Directed Combinatorial Biosynthesis of Cinnamoyl, Dihydrocinnamoyl, and Benzoyl Anthranilates in Saccharomyces cerevisiae.
abstract: 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.
authors.affiliation: Joint BioEnergy Institute, Emery Station East, 5885 Hollis St, 4th Floor, Emeryville, California, 94608, United States of America; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, 94720, United States of America.
authors.forename: Aymerick
authors.initials: A
authors.lastname: Eudes
authors.affiliation: Joint BioEnergy Institute, Emery Station East, 5885 Hollis St, 4th Floor, Emeryville, California, 94608, United States of America; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, 94720, United States of America; Graduate Program, San Francisco State University, San Francisco, California, 94132, United States of America.
authors.forename: Veronica
authors.initials: V
authors.lastname: Teixeira Benites
authors.affiliation: Joint BioEnergy Institute, Emery Station East, 5885 Hollis St, 4th Floor, Emeryville, California, 94608, United States of America; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, 94720, United States of America.
authors.forename: George
authors.initials: G
authors.lastname: Wang
authors.affiliation: Joint BioEnergy Institute, Emery Station East, 5885 Hollis St, 4th Floor, Emeryville, California, 94608, United States of America; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, 94720, United States of America.
authors.forename: Edward E K
authors.initials: EE
authors.lastname: Baidoo
authors.affiliation: Joint BioEnergy Institute, Emery Station East, 5885 Hollis St, 4th Floor, Emeryville, California, 94608, United States of America; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, 94720, United States of America.
authors.forename: Taek Soon
authors.initials: TS
authors.lastname: Lee
authors.affiliation: Joint BioEnergy Institute, Emery Station East, 5885 Hollis St, 4th Floor, Emeryville, California, 94608, United States of America; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, 94720, United States of America; Department of Bioengineering & Department of Chemical & Biomolecular Engineering, University of California, Berkeley, California, 94720, United States of America.
authors.forename: Jay D
authors.initials: JD
authors.lastname: Keasling
authors.affiliation: Joint BioEnergy Institute, Emery Station East, 5885 Hollis St, 4th Floor, Emeryville, California, 94608, United States of America; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, 94720, United States of America.
authors.forename: Dominique
authors.initials: D
authors.lastname: Loqué
mesh_terms: D020650:Combinatorial Chemistry Techniques; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D062367:ortho-Aminobenzoates / Q000378:metabolism*
chemical_list: D062367:ortho-Aminobenzoates