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In this study, Euglena gracilis wild-type … 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. iquid nitrogen followed by lyophilization.
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