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Actinorhodin-samples-2-last-few-proteins_2019-03-25_13-52-45.sky.zip2019-04-06 13:09:325882718
Actinorhodin-samples-1_2019-03-25_13-51-29.sky.zip2019-04-06 13:09:3120606327318
Programmable polyketide biosynthesis platform for production of aromatic compounds in yeast

  • Organism: Saccharomyces cerevisiae
  • Instrument: 6460 Triple Quadrupole LC/MS
  • SpikeIn: No
  • Keywords: Metabolic engineering, polyketides, S. cerevisiae, Targeted proteomics
  • Lab head: Chris Petzold
To accelerate the shift to bio-based production and overcome complicated functional implementation of natural and artificial biosynthetic pathways to industry relevant organisms, development of new, versatile, bio-based production platforms are required. Here we present a novel yeast platform for biosynthesis of bacterial aromatic polyketides. The platform is based on a synthetic polyketide synthase system enabling a first demonstration of aromatic polyketide biosynthesis in a eukaryotic host.
Experiment Description
Here, we describe a first-of-its-kind programmable polyketide production platform in yeast Saccharomyces cerevisiae, based on combining the generation of a polyketide (octaketide) by plant-based type III octaketide synthase (OKS) from Aloe arborescens to produce type II polyketide products through benzoisochromanequinone antibiotic actinorhodin pathway (Act) from Streptomyces coelicolor. To optimize the platform strain for production of type II polyketide compounds, we integrated a second copy of each of the four genes encoding ActVI-3, ActVI-2, ActVA-6, ActVB, all showing low abundances as evaluated from whole cell proteomics analysis. In an optimized strain we observed that yeast colonies were more intensely colored. To investigate if this phenotype could be correlated with increased DHK production, we performed comparative metabolite profiling by LC-MS, and noted that production of the putative DHK was indeed increased.
Created on 4/6/19, 1:09 PM

This data is available under the CC BY 4.0 license.