De novo biosynthesis of trans-cinnamic acid derivatives in Saccharomyces cerevisiae
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The production of natural aroma compounds is an expanding field within the branch of white biotechnology. Three aromatic compounds of interest are cinnamaldehyde, the typical cinnamon aroma that has applications in agriculture and medical sciences, as well as cinnamyl alcohol and hydrocinnamyl alcohol, which have applications in the cosmetic industry. Current production methods, which rely on extraction from plant materials or chemical synthesis, are associated with drawbacks regarding scalability, production time, and environmental impact. These considerations make the development of a sustainable microbial-based production highly desirable. Through steps of rational metabolic engineering, we engineered the yeast Saccharomyces cerevisiae as a microbial host to produce trans-cinnamic acid derivatives cinnamaldehyde, cinnamyl alcohol, and hydrocinnamyl alcohol, from externally added trans-cinnamic acid or de novo from glucose as a carbon source. We show that the desired products can be de novo synthesized in S. cerevisiae via the heterologous overexpression of the genes encoding phenylalanine ammonia lyase 2 from Arabidopsis thaliana (AtPAL2), aryl carboxylic acid reductase (acar) from Nocardia sp., and phosphopantetheinyl transferase (entD) from Escherichia coli, together with endogenous alcohol dehydrogenases. This study provides a proof of concept and a strain that can be further optimized for production of high-value aromatic compounds.
Keywordstrans-cinnamic acid Bioconversion Cinnamaldehyde Cinnamyl alcohol Hydrocinnamyl alcohol
Stefan Bruder is gratefully acknowledged for providing the plasmid pRS41K_optACAR_optEntD. The CGQ system was kindly provided by Aquila Biolabs, GmbH. We kindly acknowledge the support of this work by the YEASTCELL project (REA Grant No. 606795) under the EU’s Seventh Framework Programme for Research (FP7).
MG performed the experimental work, analyzed the data, and wrote the manuscript. MO and EB initiated the work on tCA production. MG, JDK, and PB designed the experiments. JDK and PB helped in data analysis. LP helped in the identification of unknown compounds. JDK, PB, MO, and EB helped in drafting the manuscript. All authors have read and approved the final manuscript.
Compliance with ethical standards
This study was funded by the European Commission under the Seventh Framework Programme, Marie-Curie ITN YEASTCELL (grant number 606795).
This article does not contain any studies with human participants or animals performed by any of the authors.
Conflict of interest
The authors declare that they have no conflict of interest.
- Bruder S, Reifenrath M, Thomik T, Boles E, Herzog K (2016) Parallelised online biomass monitoring in shake flasks enables efficient strain and carbon source dependent growth characterisation of Saccharomyces cerevisiae. Microb Cell Factories 15:127. doi: 10.1186/s12934-016-0526-3 CrossRefGoogle Scholar
- Hansen EH, Moller BL, Kock GR, Bunner CM, Kristensen C, Jensen OR, Okkels FT, Olsen CE, Motawia MS, Hansen J (2009) De novo biosynthesis of vanillin in fission yeast (Schizosaccharomyces pombe) and baker's yeast (Saccharomyces cerevisiae). Appl Environ Microbiol 75:2765–2774. doi: 10.1128/AEM.02681-08 CrossRefPubMedPubMedCentralGoogle Scholar
- Lehka BJ, Eichenberger M, Bjørn-Yoshimoto WE, Garcia Vanegas K, Buijs N, Jensen NB, Dyekjær JD, Jenssen H, Simon E, Naesby M (2017) Improving heterologous production of phenylpropanoids in Saccharomyces cerevisiae by tackling an unwanted side reaction of Tsc13, an endogenous double-bond reductase. FEMS Yeast Res 17:fox004. doi: 10.1093/femsyr/fox004 CrossRefGoogle Scholar
- Richmond HH (1947) Preparation of cinnamaldehyde. Patent no. US 2529186 AGoogle Scholar
- Sambrook J, Russel D (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
- Utchariyakiat I, Surassmo S, Jaturanpinyo M, Khuntayaporn P, Chomnawang MT (2016) Efficacy of cinnamon bark oil and cinnamaldehyde on anti-multidrug resistant Pseudomonas aeruginosa and the synergistic effects in combination with other antimicrobial agents. BMC Complement Altern Med 16:158. doi: 10.1186/s12906-016-1134-9 CrossRefPubMedPubMedCentralGoogle Scholar