Omega‐3 production by fermentation of Yarrowia lipolytica: From fed‐batch to continuous

ABSTRACT The omega‐3 fatty acid, cis‐5,8,11,14,17‐eicosapentaenoic acid (C20:5; EPA) has wide‐ranging benefits in improving heart health, immune function, and mental health. A sustainable source of EPA production through fermentation of metabolically engineered Yarrowia lipolytica has been developed. In this paper, key fed‐batch fermentation conditions were identified to achieve 25% EPA in the yeast biomass, which is so far the highest EPA titer reported in the literature. Dynamic models of the EPA fermentation process were established for analyzing, optimizing, and scaling up the fermentation process. In addition, model simulations were used to develop a two‐stage continuous process and compare to single‐stage continuous and fed‐ batch processes. The two stage continuous process, which is equipped with a smaller growth fermentor (Stage 1) and a larger production fermentor (Stage 2), was found to be a superior process to achieve high titer, rate, and yield of EPA. A two‐stage continuous fermentation experiment with Y. lipolytica strain Z7334 was designed using the model simulation and then tested in a 2 L and 5 L fermentation system for 1,008 h. Compared with the standard 2 L fed‐batch process, the two‐stage continuous fermentation process improved the overall EPA productivity by 80% and EPA concentration in the fermenter by 40% while achieving comparable EPA titer in biomass and similar conversion yield from glucose. During the long‐term experiment it was also found that the Y. lipolytica strain evolved to reduce byproduct and increase lipid production. This is one of the few continuous fermentation examples that demonstrated improved productivity and concentration of a final product with similar conversion yield compared with a fed‐batch process. This paper suggests the two‐stage continuous fermentation could be an effective process to achieve improved production of omega‐3 and other fermentation products where non‐growth or partially growth associated kinetics characterize the process. Biotechnol. Bioeng. 2017;114: 798–812. © 2016 Wiley Periodicals, Inc. With the help of an established mathematical model, a two‐stage continuous fermentation process was developed for the production of omega‐3 EPA (eicosapentaenoic acid, C20:5) by recombinant strains of Y. lipolytica. The two stage continuous experiment, which was equipped with a 2 L growth fermentor and a 5 L production fermentor, successfully improved the overall EPA productivity by 80% and EPA concentr