Abstract
This proof-of-concept study establishes Yarrowia lipolytica (Y. lipolytica) as a whole cell factory for the de novo production of long chain dicarboxylic acid (LCDCA-16 and 18) using glycerol as the sole source of carbon. Modification of the fatty acid metabolism pathway enabled creating a pool of fatty acids in a β-oxidation deficient strain. We then selectively upregulated the native fatty acid ω-oxidation pathway for the enhanced terminal oxidation of the endogenous fatty acid precursors. Nitrogen-limiting conditions and leucine supplementation were employed to induce fatty acid biosynthesis in an engineered Leu− modified strain. Our genetic engineering strategy allowed a minimum production of 330 mg/L LCDCAs in shake flask. Scale up to a 1-L bioreactor increased the titer to 3.49 g/L. Our engineered yeast also produced citric acid as a major by-product at a titer of 39.2 g/L. These results provide basis for developing Y. lipolytica as a safe biorefinery platform for the sustainable production of high-value LCDCAs from non-oily feedstock.
Highlights
Growing demand for petroleum dependent chemicals, in addition to surging environmental concerns, has inspired increased use of renewable resources
Among YlALK 2, 3, 5, 7 and 10, we noted that YlALK3, YlALK 5, and YlALK 7 each have an effect on the production of LCDCA-16 with the that YlALK3, YlALK 5, and YlALK 7 each have an effect on the production of LCDCA-16 with the following order of strength: YlALK5 > YlALK7 > YlALK3
This study reports the construction of Y. lipolytica as an oleaginous yeast cell platform for the de novo production of LCDCA monomers from renewable hydrophilic feedstock
Summary
Growing demand for petroleum dependent chemicals, in addition to surging environmental concerns, has inspired increased use of renewable resources. The sustainable production of monomers and polymers is of particular interest for reducing petroleum feedstock dependency and CO2 emissions. The environmental cost of using petrochemical polymers can be lowered by replacing them with bio-based polymers, produced through fermentation using biomass or byproducts as feedstock [1]. Glycerol is the main byproduct of biodiesel production process, and its efficient valorization would help offset biodiesel production costs. Such bioconversion decreases the environmental impact of waste streams. It is still challenging to establish an integrated bioprocess, at commercial scale, for the valorization of such hydrophilic substrates into desirable compounds [2]. The development of high productive strains and high value co-products advance the commercialization of such bioprocesses
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