Engineering Trichosporon oleaginosus for enhanced production of lipid from volatile fatty acids as carbon source

Abstract

Trichosporon oleaginosus is one of the most promising hosts for microbial lipid production owing to its high-productivity; In an effort to develop an economical production process, we engineered T. oleaginosus towards high-lipid production from volatile fatty acids (VFA) derived from anaerobic fermentation of food waste. First, we established a method for labeling intracellular lipid with lipophilic BODIPY fluorescent dye. Next, a random library was constructed by treatment with a chemical mutagen, and high-lipid producers were screened using fluorescence-activated cell sorting. Subsequently, one clone, N14, was successfully isolated, which exhibited 3-fold higher lipid production (19.4%) in VFA (6 g/L) media than the wild-type strain, and also showed increased lipid production in higher concentrations of VFA (18 or 24 g/L). Based on fatty acid methyl ester (FAME) analysis, N14 contained higher stearic acid (C18:0) and oleic acid (C18:1) content compared with those of the wild-type strain.