β-Farnesene Production from Low-Cost Glucose in Lignocellulosic Hydrolysate by Engineered Yarrowia lipolytica

beta-Farnesene is value-added acyclic volatile sesquiterpene with wide applications in energy, industry, and agriculture. Producing high-value-added compounds from low-cost renewable feedstocks in engineered microbial cell factories is an environmentally friendly and economical process for beta-farnesene biosynthesis. In this study, the potential for using engineered Yarrowia lipolytica to produce beta-farnesene from lignocellulosic hydrolysate as the carbon source was investigated. An efficient biosynthetic pathway for beta-farnesene production was established via iterative enhancement of multiple genes based on the high endogenous acetyl-CoA flux in Yarrowia lipolytica. Overexpression of mevalonate pathway genes and screening of beta-farnesene synthase resulted in a beta-farnesene titer of 245 mg L-1 in glucose media. Additional copies of mevalonate pathway genes and enhanced expression of HMG-CoA reductase and beta-farnesene synthase further increased the titer of beta-farnesene to 470 mg L. In addition, by combining metabolic engineering strategies using the lignocellulosic hydrolysate utilization strategy, the addition of Mg2+ promoted the production of beta-farnesene, and the best-performing strain produced 7.38 +/- 0.24 g L-1 beta-farnesene from lignocellulosic hydrolysate media in a 2 L fermenter after 144 h. This study shows great potential for the sustainable production of beta-farnesene from lignocellulosic biomass via engineered Yarrowia lipolytica.