De novo production of the flavonoid naringenin in engineered Saccharomyces cerevisiae

作者：Frank Koopman, Jules Beekwilder, Barbara Crimi, Adèle van Houwelingen, Robert D. Hall, Dirk Bosch, Antonius JA van Maris, Jack T. Pronk, Jean-Marc G Daran · 发表于：Microbial Cell Factories · 年份：2012 · DOI：10.1186/1475-2859-11-155 · 被引用次数：376 · 研究领域：Plant Gene Expression Analysis、Plant biochemistry and biosynthesis、Microbial Metabolic Engineering and Bioproduction

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 μM). 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 μM) in glucose-grown shake-flask cultures. In aerated, pH controlled batch reactors, extracellula...