Role of pyruvate carboxylase in accumulation of intracellular lipid of the oleaginous yeast Yarrowia lipolytica ACA-DC 50109

Abstract

Yarrowia lipolytica ACA-DC 50109 is an oleaginous yeast. In order to know the function of pyruvate carboxylase (PYC) in lipid biosynthesis, the PYC gene cloned from Pichia guilliermondii Pcla22 was overexpressed in the oleaginous yeast. The lipid contents in the wild-type strain ACA-DC 50109 and the transformants P4, P7, and P103 were 30.2 % (w/w) 36.5 % (w/w), 38.2 % (w/w), and 37.9 % (w/w). However, the amount of the secreted citric acids by strains ACA-DC 50109, P4, P77, and P103 were 0.5, 10.1, 11.5, and 9.4 g/L. In order to reduce the amount of the secreted citric acid, the PYC gene and endogenous ACL1 gene encoding ATP citrate lyase (ACL1) were simultaneously overexpressed in the oleaginous yeast. The lipid contents of the transformants PA19, PA56, PA124 were 44.4 % (w/w), 45.3 % (w/w), and 43.7 % (w/w). At the same time, the amount of the secreted citric acid by the transformants PA19, PA56, and PA124 was reduced to 5.4, 6.2, and 6.3 g/L. The PYC and ACL1 activities and their gene transcriptional levels in all the transformants were greatly enhanced compared to those in their wild-type strain ACA-DC 50109. During 10-L fermentation, lipid content in the transformant PA56 was 49.6 % (w/w) and the amount of secreted citric acid was 2.9 g/L. This meant that PYC and ACL1 can play an important role in accumulation of intracellular lipid of the oleaginous yeast Y. lipolytica ACA-DC 50109.

Systematic analysis of the lysine acetylome reveals diverse functions of lysine acetylation in the oleaginous yeast Yarrowia lipolytica

“…The compositions of the medium for lipid production were 60.0 g/L glucose, 7.0 g/L KH 2 PO 4 , 2.5 g/L Na 2 HPO 4 , 1.5 g/L MgSO 4 ·7H 2 O, 0.15 g/L CaCl 2 , 0.15 g/L FeCl 3 ·6H 2 O, 0.02 g/L ZnSO 4 ·7H 2 O, 0.06 g/L MnSO 4 ·H 2 O, and 0.5 g/L yeast extract (Wang et al 2012). The yeast strain was incubated and harvested according to the methods described (Wang et al 2015). In brief, 2.0 mL of yeast cell suspension (2.5 × 10 8 cells/mL) which had been pre-cultivated in YPD medium for 24 h were added to 50 mL of the lipid production medium.…”

“…Yarrowia lipolytica ACA-DC 50109, an oleaginous yeast, could produce lipid almost 50% (w/w) of the cell dry weight (Wang et al 2013, 2015). In this study, we performed the first investigation on the acetylproteome of this microorganism.…”

Engineering Yarrowia lipolytica for production of medium-chain fatty acids

“…Yarrowia lipolytica is the most-well studied of these yeasts, and researchers have previously engineered it for increased lipid production by a variety of rational and evolutionary approaches (Tai and Stephanopoulos 2013;Wang et al 2014;Liu et al 2015;Qiao et al 2015;Zhang et al 2014). In addition, many researchers have successfully produced novel fatty acid products in Y. lipolytica including omega-3-fatty acids, linoleic acid-derived unsaturated and polyunsaturated fatty acids, and alkanes (Xue et al 2013;Beopoulos et al 2014;Zhang et al 2013;Blazeck et al 2013;Madzak 2015).…”

Improved docosahexaenoic acid production in Aurantiochytrium by glucose limited pH-auxostat fed-batch cultivation

“…Pyruvate carboxylase (PC) has been thought to play a role in recycling of OAA as an intermediate for AcCoA and NADPH synthesis in oleaginous organisms (Ratledge, 2014). Overexpression of PC resulted in significant increases in lipid content in Y. lipolytica (Wang et al 2015). In B072, the PC activity trend during lipogenesis was comparable to that of ACL, suggesting that PC is also an important lipogenic enzyme for B072.…”