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. 2017 Nov 30;13(11):360-365.
doi: 10.6026/97320630013360. eCollection 2017.

Ymr210wp leads to the accumulation of phospholipids and steryl esters in yeast

Naresh Kumar Manda  1 , Venkata Bala Sai Chaitanya Thunuguntla  1 , Chandrasekhar Bokka  1 , Bondili Jayakumar Singh  1
Affiliations

Ymr210wp leads to the accumulation of phospholipids and steryl esters in yeast

Naresh Kumar Manda et al. Bioinformation. .

Abstract

Ymr210w was identified as a MAG (Monoacylglycerol) lipase. The accumulation of the phospholipids in the ymr210wΔ was not clearly understood. It was expressed in S. cerevisiae using pYES2/CT vector and His-tag purified recombinant protein confirmed TAG lipase activity. To further evaluate the role of YMR210w, ester hydrolase activity was also confirmed with pNP-acetate, pNP-butyrate and pNP - palmitate. GC-MS lipid profiling of ymr210wΔ showed an increase in the 15:0 Pentadecanoic acid by 76% among the total lipids. Phospholipid, Erucic acid 22:1 (Δ13) showed 43% increase while steryl esters showed significant changes with 16:0 hexadecanoic acid augmentations by 80% and 18:0 Octadecanoic acid by 165% when compared to wild type (WT). Increase in the steryl ester and TAG content supports the accumulation of lipid bodies in ymr210wΔ strain when compared with WT cells.

Keywords: Ester Hydrolase; Lipid droplets; Saccharomyces cerevisiae; Steryl esters; TAG Lipase; Triacylglycerols.

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Figures

Figure 1
Figure 1
pH and Temperature optimum: (A) Represnts pH optimum and (B) Temperature optimum for Ester hydrolase activity of ymr210wp with pNPA, pNPB and pNPP substrates. The ymr210wp has optimum activity with pH 7.5 and temperature 30°C for the substrates pNPA and pNPB. Whereas, with pNPP ymr210wp showed maximum activity at pH 8.5 and temperature 45°C.
Figure 2
Figure 2
TAG and SE lipid profiles showing variations: GC-MS analysis of FAMEs was performed on Agilent 6890N gas chromatography instrument. Lipid profiles of WT and ymr210wΔ are depicted. (A) The TAG profile shows a significant increase of 14:0, 14:1(11) lipid classes in ymr210wΔ strain compared to WT. (B) The steryl esters 16:0 and 18:0 of ymr210wΔ increased compared to WT. Whereas, 12:0, 14:1(11) and 16:1(9) lipid classes were decreased ('*' represents significant difference).
Figure 3
Figure 3
Lipid profiles estimated with GC-MS: Lipid profiles of WT and ymr210wΔ are depicted. (A) Total lipids 12:0 class significantly decreased and 16:1(9) significantly increased compare to WT. (B) Phospholipid profile of ymr210wΔ strain shows 22:1(13) increased significantly compare to WT and (C) Depicts free fatty acids of 16:1(9) significantly increased. Significance represented as '*'.
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