Print Email Facebook Twitter De novo production of the flavonoid naringenin in engineered Saccharomyces cerevisiae Title De novo production of the flavonoid naringenin in engineered Saccharomyces cerevisiae Author Koopman, F.W. Beekwilder, J. Crimi, B. Van Houwelingen, A. Hall, R.D. Bosch, D. Van Maris, A.J.A. Pronk, J.T. Daran, J.M. Faculty Applied Sciences Department BT/Biotechnology Date 2012-12-08 Abstract Background Flavonoids comprise a large family of secondary plant metabolic intermediates that exhibit a wide variety of antioxidant and human health-related properties. Plant production of flavonoids is limited by the low productivity and the complexity of the recovered flavonoids. Thus to overcome these limitations, metabolic engineering of specific pathway in microbial systems have been envisaged to produce high quantity of a single molecules. Result Saccharomyces cerevisiae was engineered to produce the key intermediate flavonoid, naringenin, solely from glucose. For this, specific naringenin biosynthesis genes from Arabidopsis thaliana were selected by comparative expression profiling and introduced in S. cerevisiae. The sole expression of these A. thaliana genes yielded low extracellular naringenin concentrations (<5.5 ?M). To optimize naringenin titers, a yeast chassis strain was developed. Synthesis of aromatic amino acids was deregulated by alleviating feedback inhibition of 3-deoxy-d-arabinose-heptulosonate-7-phosphate synthase (Aro3, Aro4) and byproduct formation was reduced by eliminating phenylpyruvate decarboxylase (Aro10, Pdc5, Pdc6). Together with an increased copy number of the chalcone synthase gene and expression of a heterologous tyrosine ammonia lyase, these modifications resulted in a 40-fold increase of extracellular naringenin titers (to approximately 200 ?M) in glucose-grown shake-flask cultures. In aerated, pH controlled batch reactors, extracellular naringenin concentrations of over 400 ?M were reached. Conclusion The results reported in this study demonstrate that S. cerevisiae is capable of de novo production of naringenin by coexpressing the naringenin production genes from A. thaliana and optimization of the flux towards the naringenin pathway. The engineered yeast naringenin production host provides a metabolic chassis for production of a wide range of flavonoids and exploration of their biological functions. Subject saccharomyces cerevisiaenaringeninde novoflavonoidsmetabolic engineeringOA-Fund TU Delft To reference this document use: http://resolver.tudelft.nl/uuid:85a25098-e9d3-4262-931e-260e27a7ab82 DOI https://doi.org/10.1186/1475-2859-11-155 Publisher BioMed Central ISSN 1475-2859 Source http://www.microbialcellfactories.com/content/11/1/155 Source Microbial Cell Factories, 11, 2012 Part of collection Institutional Repository Document type journal article Rights © 2012 The Author(s)Licensee BioMed Central Ltd. - This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited Files PDF Koopman_2012.pdf 673.94 KB Close viewer /islandora/object/uuid:85a25098-e9d3-4262-931e-260e27a7ab82/datastream/OBJ/view