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

Supplementary data associated with this article can be found, inthe online version, at http://dx.doi.org/10.1016/j.bej.2017.05.015.
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 2.5 mM; caffeic acid production reached 370 M. Regarding curcumin, 17 M 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.
FundingThis study was supported by the Portuguese Foundationfor Science and Technology (FCT) under the scope of thestrategic funding of UID/BIO/04469/2013 unit and COMPETE2020 (POCI-01-0145-FEDER-006684) and under the scope ofthe Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462). The authors also acknowledge financial support fromBioTecNorte operation (NORTE-01-0145-FEDER-000004) and theproject MultiBiorefinery (POCI-01-0145-FEDER-016403) funded by the European Regional Development Fund under the scope ofNorte2020 − Programa Operacional Regional do Norte and the Post-doctoral grant (UMINHO/BPD/37/2015) to J. L. Rodrigues funded byFCT.
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