Cell wall canals formed upon growth of Candida maltosa in the presence of hexadecane are associated with polyphosphates.

Canals are supramolecular complexes observed in the cell wall of Candida maltosa grown in the presence of hexadecane as a sole carbon source. Such structures were not observed in glucose-grown cells. Microscopic observations of cells stained with diaminobenzidine revealed the presence of oxidative enzymes in the canals. 4΄,6΄-diamino-2-phenylindole staining revealed that a substantial part of cellular polyphosphate was present in the cell wall of cells grown on hexadecane in condition of phosphate limitation. The content and chain length of polyphosphates were higher in hexadecane-grown cells than in glucose grown ones. The treatment of cells with yeast polyphosphatase PPX1 resulted in the decrease of the canal size. These data clearly indicated that polyphosphates are constituents of canals; they might play an important role in the canal structure and functioning.

Modifications of the cell wall of yeasts grown on hexadecane and under starvation conditions.

Electron-microscopic examinations have demonstrated local modifications in the cell wall of the yeast Candida maltosa grown on hexadecane. In our earlier studies, these modified sites, observed in other yeasts grown on oil hydrocarbons, were conventionally called 'canals'. The biochemical and cytochemical studies of C. maltosa have revealed a correlation between the formation of 'canals' and decrease in the amount of cell wall polysaccharides, glucan and mannan. The ultrathin sections and surface replicas have shown that the 'canals' are destroyed by pronase, thus indicating that a significant proportion of their content is represented by proteins. This finding was compatible with our earlier data on the localization of oxidative enzymes in 'canals' and possible participation of the 'canals' in the primary oxidation of hydrocarbons. A completely unexpected and intriguing phenomenon has been the appearance of 'canals' in the yeast C. maltosa under starvation conditions. Unlike the yeasts grown on hexadecane, mannan almost disappears in starving cells, while the quantity of glucan first decreases and then is restored to its initial level. The role of 'canals' in starving cells is as yet unclear; it is assumed that they acquire exoenzymes involved in the utilization of products of cell lysis in the starving population. In the future, 'canals' of starving cells will be studied in connection with their possible participation in apoptosis.

Microorganisms form exocellular structures, trophosomes, to facilitate biodegradation of oil in aqueous media.

Cytochemical staining and microscopy were used to study the trophic structures and cellular morphotypes that are produced during the colonization of oil-water interfaces by oil-degrading yeasts and bacteria. Among the microorganisms studied here, the yeasts (Schwanniomyces occidentalis, Torulopsis candida, Candida tropicalis, Candida lipolytica, Candida maltosa, Candida paralipolytica) and two representative bacteria (Rhodococcus sp. and Pseudomonas putida) produced exocellular structures composed of biopolymers during growth on petroleum hydrocarbons. Four of the yeasts including S. occidentalis, T. candida, C. tropicalis and C. maltosa excreted polymers through modified sites in their cell wall ('canals'), whereas C. lipolytica and C. paralipolytica and the two bacterial species secreted polymers over the entire cell surface. These polymers took the form of fibrils and films that clogged pores and cavities on the surfaces of the oil droplets. A three-dimensional reconstruction of the cavities using serial thin sections showed that the exopolymer films isolated the ambient aqueous medium together with microbial cells and oil to form both closed and open granules that contained pools of oxidative enzymes utilized for the degradation of the oil hydrocarbons. The formation of such granules, or 'trophosomes,' appears to be a fundamental process that facilitates the efficient degradation of oil in aqueous media.