Highly flexible metabolism of the marine euglenozoan protist Diplonema papillatum

Background The phylum Euglenozoa is a group of flagellated protists comprising the diplonemids, euglenids, symbiontids, and kinetoplastids. The diplonemids are highly abundant and speciose, and recent tools have rendered the best studied representative, Diplonema papillatum, genetically tractable. However, despite the high diversity of diplonemids, their lifestyles, ecological functions, and even primary energy source are mostly unknown. Results We designed a metabolic map of D. papillatum cellular bioenergetic pathways based on the alterations of transcriptomic, proteomic, and metabolomic profiles obtained from cells grown under different conditions. Comparative analysis in the nutrient-rich and nutrient-poor media, as well as the absence and presence of oxygen, revealed its capacity for extensive metabolic reprogramming that occurs predominantly on the proteomic rather than the transcriptomic level. D. papillatum is equipped with fundamental metabolic routes such as glycolysis, gluconeogenesis, TCA cycle, pentose phosphate pathway, respiratory complexes, β-oxidation, and synthesis of fatty acids. Gluconeogenesis is uniquely dominant over glycolysis under all surveyed conditions, while the TCA cycle represents an eclectic combination of standard and unusual enzymes. Conclusions The identification of conventional anaerobic enzymes reflects the ability of this protist to survive in low-oxygen environments. Furthermore, its metabolism quickly reacts to restricted carbon availability, suggesting a high metabolic flexibility of diplonemids, which is further reflected in cell morphology and motility, correlating well with their extreme ecological valence.

Accelerated Alcoholic Fermentation of Intact Grapes by Saccharomyces Cerevisiae in Symbiosis with Microbial Community Inhabiting Grape-skin

Saccharomyces cerevisiae, an essential player in alcoholic fermentation during winemaking, is rarely found in intact grapes. Here, we addressed symbiotic interactions between S. cerevisiae and grape-skin residents upon spontaneous wine fermentation. When glucose was used as a carbon source, the yeast-like fungus Aureobasidium pullulans, a major grape-skin resident, had no effect on alcoholic fermentation by S. cerevisiae. In contrast, when intact grape berries as a sole carbon source, coculture of S. cerevisiae and A. pullulans accelerated alcoholic fermentation. Thus, grape-inhabiting microorganisms may increase carbon availability by degrading and/or incorporating grape-skin materials, such as cell wall and cuticles. A. pullulans exhibited broad spectrum assimilation of plant-derived carbon sources, including ω-hydroxy fatty acids, arising from degradation of cutin. In fact, yeast-type cutinase was produced from A. pullulans EXF-150 strain. The degradation and utilization of grape-skin materials by fungal microbiota may account for their colonization on grape-skin and symbiotic interactions with S. cerevisiae.

Passive plasma membrane transporters play a critical role in perception of carbon availability in yeast

Recently, our lab found that the canonical glucose/galactose regulation pathway in yeast makes the decision to metabolize galactose based on the ratio of glucose to galactose concentrations in the external medium. This led to the question of where and how the ratio-sensing is achieved. Here, we consider the possibilities of an intracellular, extracellular, or membrane bound ratio sensing mechanisms. We show that hexose transporters in the plasma membrane are mainly responsible for glucose/galactose ratio-sensing in yeast. Further, while the glucose sensors Gpr1, Snf3, and Rgt2 are not required for ratio sensing, they help modulate the ratio sensing phenotype by regulating the expression of individual transporters in different environments. Our study provides an example of an unexpected, but potentially widespread, mechanism for making essential decisions.