De novo production of the flavonoid naringenin in engineered Saccharomyces cerevisiae View Full Text


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Article Info

DATE

2012-12-08

AUTHORS

Frank Koopman, Jules Beekwilder, Barbara Crimi, Adele van Houwelingen, Robert D Hall, Dirk Bosch, Antonius JA van Maris, Jack T Pronk, Jean-Marc Daran

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 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, extracellular naringenin concentrations of over 400 μM 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. More... »

PAGES

155-155

References to SciGraph publications

  • 2009-03-05. Overexpression of Acetyl-CoA Carboxylase Gene of Mucor rouxii Enhanced Fatty Acid Content in Hansenula polymorpha in MOLECULAR BIOTECHNOLOGY
  • 2007-01-31. High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method in NATURE PROTOCOLS
  • 2009-07-01. Biosynthesis and biotechnological production of flavanones: current state and perspectives in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2009-08-02. Synthetic protein scaffolds provide modular control over metabolic flux in NATURE BIOTECHNOLOGY
  • 1998-05. The two 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase isoenzymes from Saccharomyces cerevisiae show different kinetic modes of inhibition in ARCHIVES OF MICROBIOLOGY
  • 2012. Functional expression and characterization of five wax ester synthases in Saccharomyces cerevisiae and their utility for biodiesel production in BIOTECHNOLOGY FOR BIOFUELS
  • 2011-07-06. Metabolic engineering of flavonoids in plants and microorganisms in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2012-02-24. Functional expression and characterization of five wax ester synthases in Saccharomyces cerevisiae and their utility for biodiesel production in BIOTECHNOLOGY FOR BIOFUELS
  • 2009-08-13. Yeast artificial chromosomes employed for random assembly of biosynthetic pathways and production of diverse compounds in Saccharomyces cerevisiae in MICROBIAL CELL FACTORIES
  • 2007-04-05. Untargeted large-scale plant metabolomics using liquid chromatography coupled to mass spectrometry in NATURE PROTOCOLS
  • 2012-03-26. De novo sequencing, assembly and analysis of the genome of the laboratory strain Saccharomyces cerevisiae CEN.PK113-7D, a model for modern industrial biotechnology in MICROBIAL CELL FACTORIES
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1186/1475-2859-11-155

    DOI

    http://dx.doi.org/10.1186/1475-2859-11-155

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1004817875

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/23216753


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    28 schema:description 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, extracellular naringenin concentrations of over 400 μM 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.
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    38 acid
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