Abstract
BackgroundYarrowia lipolytica, a non-traditional oil yeast, has been widely used as a platform for lipid production. However, the production of other chemicals such as terpenoids in engineered Y. lipolytica is still low. α-Farnesene, a sesquiterpene, can be used in medicine, bioenergy and other fields, and has very high economic value. Here, we used α-farnesene as an example to explore the potential of Y. lipolytica for terpenoid production.ResultsWe constructed libraries of strains overexpressing mevalonate pathway and α-farnesene synthase genes by non-homologous end-joining (NHEJ) mediated integration into the Y. lipolytica chromosome. First, a mevalonate overproduction strain was selected by overexpressing relevant genes and changing the cofactor specificity. Based on this strain, the downstream α-farnesene synthesis pathway was overexpressed by iterative integration. Culture conditions were also optimized. A strain that produced 25.55 g/L α-farnesene was obtained. This is the highest terpenoid titer reported in Y. lipolytica.ConclusionsYarrowia lipolytica is a potentially valuable species for terpenoid production, and NHEJ-mediated modular integration is effective for expression library construction and screening of high-producer strains.
Highlights
Yarrowia lipolytica, a non-traditional oil yeast, has been widely used as a platform for lipid production
Construction of high mevalonate production strains through non-homologous end-joining (NHEJ)‐mediated genome integration α-Farnesene is synthesized using farnesyl pyrophosphate (FPP) as precursor, which is the intermediate product of the MVA pathway
MVA is synthesized from acetyl-CoA through the action of three enzymes—acetyl-CoA acetyltransferase (ERG10/ AtoB), HMG-CoA synthase (ERG13/HMGS), and HMGCoA reductase (HMGR) (Fig. 1)
Summary
A non-traditional oil yeast, has been widely used as a platform for lipid production. As the oleaginous yeast, is emerging as a model non-conventional oleaginous yeast [1]. It has been widely recognized as a valuable host for the production of lipid-based biofuels and oleo chemicals and has “generally regarded as safe” (GRAS) status [2, 3]. Markham et al demonstrated that Y. lipolytica can produce a type III polyketide triacetic acid lactone (TAL) with a titer of 35.9 g/L, and a previously uncharacterized pyruvate bypass pathway was identified as a key pathway to increase TAL production [9].
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