title: De novo biosynthesis of trans-cinnamic acid derivatives in Saccharomyces cerevisiae.
abstract: 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.
authors.affiliation: Institute of Molecular Biosciences, Goethe University Frankfurt, Max-von-Laue Straße 9, 60438, Frankfurt am Main, Germany.
authors.forename: Manuela
authors.initials: M
authors.lastname: Gottardi
authors.affiliation: Department of Biotechnology and Biosciences, University of Milano-Bicocca, P.zza della Scienza 4, 20126, Milan, Italy.|Terranol A/S, c/o Section for Sustainable Biotechnology, Aalborg University, Copenhagen A.C. Meyers Vænge 15, DK-2450, Copenhagen, SV, Denmark.
authors.forename: Jan Dines
authors.initials: JD
authors.lastname: Knudsen
authors.affiliation: Metabolic Explorer, Biopôle Clermont Limagne, 63360, Saint-Beauzire, France.
authors.forename: Lydie
authors.initials: L
authors.lastname: Prado
authors.affiliation: Institute of Molecular Biosciences, Goethe University Frankfurt, Max-von-Laue Straße 9, 60438, Frankfurt am Main, Germany. m.oreb@uni.bio-frankfurt.de.
authors.forename: Mislav
authors.identifier: http://orcid.org/0000-0002-6118-1517
authors.initials: M
authors.lastname: Oreb
authors.affiliation: Department of Biotechnology and Biosciences, University of Milano-Bicocca, P.zza della Scienza 4, 20126, Milan, Italy. paola.branduardi@unimib.it.
authors.forename: Paola
authors.initials: P
authors.lastname: Branduardi
authors.affiliation: Institute of Molecular Biosciences, Goethe University Frankfurt, Max-von-Laue Straße 9, 60438, Frankfurt am Main, Germany.
authors.forename: Eckhard
authors.initials: E
authors.lastname: Boles
mesh_terms: D000171:Acrolein / Q000031:analogs & derivatives / Q000378:metabolism; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D053898:Biosynthetic Pathways; D002934:Cinnamates / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D009615:Nocardia / Q000201:enzymology / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics; D010650:Phenylalanine Ammonia-Lyase / Q000235:genetics; D000075082:Proof of Concept Study; D020005:Propanols / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
chemical_list: D002934:Cinnamates; D020005:Propanols; C029010:cinnamic acid; D000171:Acrolein; D010088:Oxidoreductases; C019825:carboxylic acid reductase; D010650:Phenylalanine Ammonia-Lyase; D005947:Glucose; C012843:cinnamaldehyde; C020722:cinnamyl alcohol
keywords: Bioconversion; Cinnamaldehyde; Cinnamyl alcohol; Hydrocinnamyl alcohol; trans-cinnamic acid