Critical Involvement of Environmental Carbon Dioxide Fixation to Drive Wax Ester Fermentation in Euglena

• Published in: PloS one 2016-09, Vol.11 (9), p.e0162827-e0162827
• Main Authors: Padermshoke, Adchara, Ogawa, Takumi, Nishio, Kazuki, Nakazawa, Masami, Nakamoto, Masatoshi, Okazawa, Atsushi, Kanaya, Shigehiko, Arita, Masanori, Ohta, Daisaku
• Format: Article
• Language: English
• Subjects: Alcohol; Anoxia; Assimilation; Biology and Life Sciences; Cancer; Carbon 13; Carbon dioxide; Carbon dioxide fixation; Carbon sources; Chromatography; Citric acid; Enrichment; Environmental conditions; Environmental science; Esters; Euglena; Euglena gracilis; Fatty acids; Fermentation; Hypoxia; Information science; Inorganic carbon; Intermediates; Isotopic enrichment; Malate; Mass spectrometry; Medicine and Health Sciences; Metabolism; Metabolites; Microorganisms; Physical Sciences; Research and Analysis Methods; Scientific imaging; Tricarboxylic acid cycle
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0162827

Accumulation profiles of wax esters in Euglena gracilis Z were studied under several environmental conditions. The highest amount of total wax esters accumulated under hypoxia in the dark, and C28 (myristyl-myristate, C14:0-C14:0) was prevalent among all conditions investigated. The wax ester production was almost completely suppressed under anoxia in the light, and supplying exogenous inorganic carbon sources restored wax ester fermentation, indicating the need for external carbon sources for the wax ester fermentation. 13C-labeling experiments revealed specific isotopic enrichment in the odd-numbered fatty acids derived from wax esters, indicating that the exogenously-supplied CO2 was incorporated into wax esters via the propionyl-CoA pathway through the reverse tricarboxylic acid (TCA) cycle. The addition of 3-mercaptopicolinic acid, a phosphoenolpyruvate carboxykinase (PEPCK) inhibitor, significantly affected the incorporation of 13C into citrate and malate as the biosynthetic intermediates of the odd-numbered fatty acids, suggesting the involvement of PEPCK reaction to drive wax ester fermentation. Additionally, the 13C-enrichment pattern of succinate suggested that the CO2 assimilation might proceed through alternative pathways in addition to the PEPCK reaction. The current results indicate that the mechanisms of anoxic CO2 assimilation are an important target to reinforce wax ester fermentation in Euglena.