α-Farnesene production from lipid by engineered Yarrowia lipolytica

Abstract

Producing high value-added products from waste lipid feedstock by microbial cell factory has great advantages to minimize the pollution as well as improve the economic value of wasted oils and fats. Yarrowia lipolytica is a non-conventional oleaginous yeast and can grow on a variety of hydrophobic substrates. In this study, we explored its ability to synthesize α-farnesene, an important sesquiterpene, using lipid feedstock. Based on the α-farnesene production strain, we constructed previously, we identified that Erg12 was the key limiting factor to further increase the α-farnesene production. The α-farnesene production was improved by 35.8% through increasing the copy number of ERG12 and FSERG20 on oleic acid substrate. Expression of heterologous VHb further improved α-farnesene production by 12.7%. Combining metabolic engineering with the optimization of fermentation conditions, the α-farnesene titer and yield reached 10.2 g/L and 0.1 g/g oleic acid, respectively, in fed-batch cultivation. The α-farnesene synthesis ability on waste cooking oil and other edible oils were also explored. Compared with using glucose as carbon source, using lipid substrates obtained higher α-farnesene yield and titer, but lower by-products accumulation, demonstrating the advantage of Y. lipolytica to synthesize high value-added products using lipid feedstock.

Engineering Yarrowia lipolytica for the sustainable production of β-farnesene from waste oil feedstock

“…The accumulations of lipids and squalene were 102.9 and 1.71 mg/g DCW, respectively. The accumulations of these by-products were lower than that when using glucose as the carbon source [ 12 , 13 ].

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“…Our recent work indicated that Y. lipolytica is also a potential farnesene production strain, and α-farnesene could be produced at 25.5 g/L [ 12 ]. However, all the strategies described above used sugar as the feedstock [ 11 , 12 ], and there are few studies on the use of lipid feedstock to produce farnesene [ 13 ].…”

Cofactor Engineering for Efficient Production of α-Farnesene by Rational Modification of NADPH and ATP Regeneration Pathway in Pichia pastoris

“…α-Farnesene is biosynthesized from farnesyl pyrophosphate (FPP) and dimethylallyl pyrophosphate (DMAPP), which are mainly produced through the mevalonate (MVA) pathway in P. pastoris [ 3 , 6 ], and thus six molecules of NADPH and nine molecules of ATP are needed to produce one molecule of α-farnesene ( Figure 1 ). Although many effective strategies have been used to increase α-farnesene production in yeast, for example, in P. pastoris [ 3 ], Saccharomyces cerevisiae [ 50 , 51 ] and Yarrowia lipolytica [ 2 , 52 ], these studies mainly focused on modifying the carbon’s metabolism pathway to enhance the carbon flux in the α-farnesene biosynthetic pathway. We first attempted to rationally reconstruct the NADPH and ATP biosynthetic pathways in order to increase α-farnesene production in an α-farnesene-producing strain, P. pastoris X33-30*.…”

Cofactor engineering for efficient production of α-farnesene by rational modification of NADPH and ATP regeneration pathway in Pichia pastoris

“…1). Although many effective strategies have been used to increase the α-farnesene production in yeast, for example, in P. pastoris 3 ,Saccharomyces cerevisiae [51][52] andYarrowia lipolytica [53][54] , these studies mainly focused on modifying the carbon's metabolism pathway to enhance the carbon flux in α-farnesene biosynthetic pathway. Here, we first tried to rationally reconstruct the biosynthetic pathways of NADPH and ATP to further increase the α-farnesene production in a α-farnesene high-producing strain P. pastoris X33-30*.…”