SE184:/S1

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Sample Set Information

ID TSE1342
Title Wax ester and lipophilic compound profiling of Euglena gracilis by gas chromatography-mass spectrometry: toward understanding of wax ester fermentation under hypoxia
Description Lipids are being increasingly used as biodiesel feedstock, and several saturated wax esters from Euglena gracilis are candidates for outdoor bulk production. Wax ester fermentation in Euglena is strongly increased by hypoxia, but key events underlying the metabolic shift toward wax ester biosynthesis are poorly understood. Profiling of wax esters and other lipophilic compounds is the first step for research toward the clarification of wax ester fermentation molecular mechanisms, and thus, a simple and comprehensive platform for their profiling is required. In this study, we established a profiling method for wax esters and lipophilic compounds in Euglena using gas chromatography-mass spectrometry (GC–MS). Using this method, we compared accumulation profiles of wax esters and lipophilic compounds between a wild-type Euglena Z strain and a bleached SM-ZK strain. Both the wild-type and the bleached strains contained C14:0 fatty acid-C14:0 fatty alcohol as a dominant wax ester. Wax ester fermentation initiated 4 h after the cessation of oxygen supply by halting the culture agitation resulting in linear increase and proportional changes of wax ester amounts during 24 h. However, complete anoxia by nitrogen gas aeration inhibited wax ester production and the addition of bicarbonates reversed the inhibition, suggesting that there is a need for an additional carbon source for wax ester fermentation under anoxia. Our simple method enables the investigation of metabolic responses leading to wax ester fermentation in Euglena.
Authors Furuhashi, T., Ogawa, T., Nakai, R., Nakazawa, M., Okazawa, A., Padermschoke, A., Nishio, K., Hirai, M.Y., Arita, M. and Ohta, D.
Reference Metabolomics, February 2015, Volume 11, Issue 1, pp 175–183
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Sample Information

ID S1
Title Euglena gracilis
Organism - Scientific Name Euglena gracilis
Organism - ID NCBI taxonomy 3039
Compound - ID
Compound - Source
Preparation In this study, Euglena gracilis wild-type Z strain and a streptomycin-bleached mutant SM-ZK with less chloroplasts (Mccalla, 1963; Oda et al., 1982) were used. They were maintained in 150 mL of Koren-Hutner medium (pH 5.0) in Sakaguchi flasks at 27 °C under continuous light on a shaker (120 rpm) for 2 days (early-log phase) or 4 days (late-log phase) (Koren and Hutner 1967). Wax ester fermentation was induced either by halting the culture agitation, which stops aeration and induces hypoxia, or by aerating N2 gas into culture medium for 10 min to remove oxygen and other gases, including carbon dioxide, completely. For anoxic conditions induced by N2 gas aeration, different NaHCO3 concentration (1 and 10 mM) were added to investigate the effect of a carbon source on wax fermentation. Cells were collected 0, 4, and 24 h after the initiation of anaerobiosis and were immediately immersed in liquid nitrogen followed by lyophilization.
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