Impact of central carbon metabolism bypasses on the production of beta-carotene inYarrowia lipolytica

Author:

Markuš TadejORCID,Soldat Mladen,Magdevska Vasilka,Horvat Jaka,Kavšček MartinORCID,Kosec Gregor,Fujs Štefan,Petrovič UrošORCID

Abstract

AbstractYarrowia lipolyticais an oleaginous yeast with ever growing popularity in the metabolic engineering circles. It is well known for its ability to accommodate a high carbon flux through acetyl-CoA and is being extensively studied for production of chemicals derived from it.We investigated the effects of modifying the upstream metabolism leading to acetyl-CoA on beta-carotene production, including its titer, yield, and content. We examined the pyruvate and the phosphoketolase bypass, both of which are stoichiometrically favorable for the production of acetyl-CoA and beta-carotene. Additionally, we examined a set of genes involved in the carnitine shuttle. We constructed a set of parental strains derived from theY. lipolyticaYB-392 wild-type strain, each with a different capacity for beta-carotene production, and introduced genes for the metabolic bypasses in each of the constructed parental strains. Subsequently, we subjected these constructed strains to a series of fermentation experiments.We discovered that altering the upstream metabolism in most cases led to a decrease in performance for production of beta-carotene. Most notably, a set of genes used for the pyruvate bypass (YlPDC2,YlALD5, andYlACS1) and the phosphoketolase bypass (LmXPKandCkPTA) resulted in the reduction of more than 30%.Our findings contribute to our understanding ofY. lipolytica’s metabolic capacity and suggest that production of beta-carotene is most likely not limited solely by the acetyl-CoA supply. We also highlight a complex nature of engineeringY. lipolytica, as most of the results from studies using a different strain background did not align with our findings.

Publisher

Cold Spring Harbor Laboratory

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