Production of tranilast [N-(3',4'-dimethoxycinnamoyl)-anthranilic acid] and its analogs in yeast Saccharomyces cerevisiae.
Biological synthesis of therapeutic drugs beneficial for human health using microbes offers an alternative production strategy to the methods that are commonly employed such as direct extraction from source organisms or chemical synthesis. In this study, we evaluated the potential for yeast (Saccharomyces cerevisiae) to be used as a catalyst for the synthesis of tranilast and various tranilast analogs (cinnamoyl anthranilates). Several studies have demonstrated that these phenolic amides have antioxidant properties and potential therapeutic benefits including antiinflammatory, antiproliferative, and antigenotoxic effects. The few cinnamoyl anthranilates naturally produced in plants such as oats and carnations result from the coupling of various hydroxycinnamoyl-CoAs to anthranilic acid. In order to achieve the microbial production of tranilast and several of its analogs, we engineered a yeast strain to co-express a 4-coumarate/CoA ligase (4CL, EC 6.2.1.12) from Arabidopsis thaliana and a hydroxycinnamoyl/benzoyl-CoA/anthranilate N-hydroxycinnamoyl/benzoyltransferase (HCBT, EC 2.3.1.144) from Dianthus caryophyllus. This modified yeast strain allowed us to produce tranilast and 26 different cinnamoyl anthranilate molecules within a few hours after exogenous supply of various combinations of cinnamic acids and anthranilate derivatives. Our data demonstrate the feasibility of rapidly producing a wide range of defined cinnamoyl anthranilates in yeast and underline a potential for the biological designed synthesis of naturally and non-naturally occurring molecules.
Eudes
Aymerick
A
Joint BioEnergy Institute, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720, USA.
Baidoo
Edward E K
EE
Yang
Fan
F
Burd
Helcio
H
Hadi
Masood Z
MZ
Collins
F William
FW
Keasling
Jay D
JD
Loqué
Dominique
D
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism*; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D001709:Biotechnology / Q000379:methods*; D029749:Dianthus / Q000201:enzymology / Q000235:genetics; D004345:Drug Industry / Q000379:methods*; D005818:Genetic Engineering; D006801:Humans; D053858:Metabolic Networks and Pathways / Q000235:genetics; D030781:Organisms, Genetically Modified; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D062367:ortho-Aminobenzoates / Q000378:metabolism*
D000894:Anti-Inflammatory Agents, Non-Steroidal; D010940:Plant Proteins; D062367:ortho-Aminobenzoates; C012293:tranilast