Differential analysis of lipids in xylose fermenting S. cerevisiae 424A (LNH-ST) and its ethanol resistant variant S. cerevisiae 424A (LNH-ST)-ER

Jin Hee Kim, Jiri Adamec, Nathan S. Mosier, Nancy W.Y. Ho, Miroslav Sedlak

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Through selective culturing/adaptation we have developed a new strain of Saccharomyces cerevisiae 424A(LNH-ST) with improved xylose fermentation and ethanol resistance compared to the original strain. The new strain has 500% higher ethanol volumetric productivity on xylose in the presence of higher ethanol concentrations (above 6% w/v) than the original strain. Since membrane lipid composition is expected to be one of the core factors to increased ethanol resistance, it is important to know the dynamics of the lipidome to understand molecular basis of the improved resistance. We analyzed lipids compositions in our original strain 424A(LNH-ST) and the new 424A(LNH-ST)-ER strain during xylose fermentation with and without ethanol addition. The concentrations of 16 free fatty acids (FFAs) were quantified by using QQQ LC/MS/MS mass spectrometry. Based on the results, stearic acid and palmitic acid were the predominant FFAs present in yeasts under the tested conditions. Interestingly, the saturated /unsaturated ratios of the FFAs were significantly different between the two strains. The 424A(LNH-ST)-ER strain contained higher concentrations of saturated FFAs than the original strain and the ratio of saturated/unsaturated FFAs decreased as ethanol concentration increased. The ratios and amounts of glycerophospholipids and sphingolipids in the cell membrane were also investigated. We have established a new approach for comparative and semi-quantitative lipidomics using RPLC (C8) coupled to LTQ-Orbitrap mass spectrometry. After data normalization of four replicate experiments, ∼70 lipid species were identified based on their molecular signatures, in which ∼30% of the species showed at least 2 fold change with a significance p>0.05 between the control and the enhanced ethanol resistant strain.

Original languageEnglish (US)
Title of host publication11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings
StatePublished - Dec 1 2011
Event2011 AIChE Annual Meeting, 11AIChE - Minneapolis, MN, United States
Duration: Oct 16 2011Oct 21 2011

Publication series

Name11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings

Conference

Conference2011 AIChE Annual Meeting, 11AIChE
CountryUnited States
CityMinneapolis, MN
Period10/16/1110/21/11

Fingerprint

Xylose
Lipids
Ethanol
Fatty acids
Nonesterified Fatty Acids
Yeast
Fermentation
Mass spectrometry
Glycerophospholipids
Palmitic acid
Sphingolipids
Stearic acid
Palmitic Acid
Cell membranes
Membrane Lipids
Chemical analysis
Productivity

ASJC Scopus subject areas

  • Chemical Engineering(all)

Cite this

Kim, J. H., Adamec, J., Mosier, N. S., Ho, N. W. Y., & Sedlak, M. (2011). Differential analysis of lipids in xylose fermenting S. cerevisiae 424A (LNH-ST) and its ethanol resistant variant S. cerevisiae 424A (LNH-ST)-ER. In 11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings (11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings).

Differential analysis of lipids in xylose fermenting S. cerevisiae 424A (LNH-ST) and its ethanol resistant variant S. cerevisiae 424A (LNH-ST)-ER. / Kim, Jin Hee; Adamec, Jiri; Mosier, Nathan S.; Ho, Nancy W.Y.; Sedlak, Miroslav.

11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings. 2011. (11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings).

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Kim, JH, Adamec, J, Mosier, NS, Ho, NWY & Sedlak, M 2011, Differential analysis of lipids in xylose fermenting S. cerevisiae 424A (LNH-ST) and its ethanol resistant variant S. cerevisiae 424A (LNH-ST)-ER. in 11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings. 11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings, 2011 AIChE Annual Meeting, 11AIChE, Minneapolis, MN, United States, 10/16/11.
Kim JH, Adamec J, Mosier NS, Ho NWY, Sedlak M. Differential analysis of lipids in xylose fermenting S. cerevisiae 424A (LNH-ST) and its ethanol resistant variant S. cerevisiae 424A (LNH-ST)-ER. In 11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings. 2011. (11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings).
Kim, Jin Hee ; Adamec, Jiri ; Mosier, Nathan S. ; Ho, Nancy W.Y. ; Sedlak, Miroslav. / Differential analysis of lipids in xylose fermenting S. cerevisiae 424A (LNH-ST) and its ethanol resistant variant S. cerevisiae 424A (LNH-ST)-ER. 11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings. 2011. (11AIChE - 2011 AIChE Annual Meeting, Conference Proceedings).
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abstract = "Through selective culturing/adaptation we have developed a new strain of Saccharomyces cerevisiae 424A(LNH-ST) with improved xylose fermentation and ethanol resistance compared to the original strain. The new strain has 500{\%} higher ethanol volumetric productivity on xylose in the presence of higher ethanol concentrations (above 6{\%} w/v) than the original strain. Since membrane lipid composition is expected to be one of the core factors to increased ethanol resistance, it is important to know the dynamics of the lipidome to understand molecular basis of the improved resistance. We analyzed lipids compositions in our original strain 424A(LNH-ST) and the new 424A(LNH-ST)-ER strain during xylose fermentation with and without ethanol addition. The concentrations of 16 free fatty acids (FFAs) were quantified by using QQQ LC/MS/MS mass spectrometry. Based on the results, stearic acid and palmitic acid were the predominant FFAs present in yeasts under the tested conditions. Interestingly, the saturated /unsaturated ratios of the FFAs were significantly different between the two strains. The 424A(LNH-ST)-ER strain contained higher concentrations of saturated FFAs than the original strain and the ratio of saturated/unsaturated FFAs decreased as ethanol concentration increased. The ratios and amounts of glycerophospholipids and sphingolipids in the cell membrane were also investigated. We have established a new approach for comparative and semi-quantitative lipidomics using RPLC (C8) coupled to LTQ-Orbitrap mass spectrometry. After data normalization of four replicate experiments, ∼70 lipid species were identified based on their molecular signatures, in which ∼30{\%} of the species showed at least 2 fold change with a significance p>0.05 between the control and the enhanced ethanol resistant strain.",
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