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Hydroxycinnamic acids and curcumin production in engineered Escherichia coli using heat shock promoters

Rodrigues, Joana L., Couto, Márcia R., Araújo, Rafael G., Prather, Kristala L.J., Kluskens, Leon, Rodrigues, Lígia R.
Biochemical engineering journal 2017 v.125 pp. 41-49
Arabidopsis thaliana, Curcuma longa, Escherichia coli, Rhodopseudomonas palustris, Rhodotorula glutinis, Saccharothrix espanaensis, ammonia, antineoplastic activity, binding sites, biochemical pathways, caffeic acid, coumarate-CoA ligase, curcumin, cytochrome P-450, genes, heat, p-coumaric acid, promoter regions, ribosomes, translation (genetics), tyrosine
Hydroxycinnamic acids and curcumin are compounds with great therapeutic potential, including anticancer properties. In this study, p-coumaric acid, caffeic acid and curcumin were produced in Escherichia coli. Their production was induced by heat using the dnaK and ibpA heat shock promoters. The ribosome binding site (RBS) used was tested and further optimized for each gene to assure an efficient translation. p-Coumaric acid was successfully produced from tyrosine and caffeic acid was produced either from tyrosine or p-coumaric acid using tyrosine ammonia lyase (TAL) from Rhodotorula glutinis, 4-coumarate 3-hydroxylase (C3H) from Saccharothrix espanaensis or cytochrome P450 CYP199A2 from Rhodopseudomonas palustris. The highest p-coumaric acid production obtained was 2.5mM; caffeic acid production reached 370μM. Regarding curcumin, 17μM was produced using 4-coumarate-CoA ligase (4CL1) from Arabidopsis thaliana, diketide-CoA synthase (DCS) and curcumin synthase 1 (CURS1) from Curcuma longa. Stronger RBSs and/or different induction conditions should be further evaluated to optimize those production levels. Herein it was demonstrated that the biosynthetic pathway of p-coumaric acid, caffeic acid and curcumin in E. coli can be triggered by using heat shock promoters, suggesting its potential for the development of new industrial bioprocesses or even new bacterial therapies.