Stela Križanović

Characterization of a S-adenosyl-l-methionine (SAM)-accumulating strain of Scheffersomyces stipitis.

S-adenosyl-l-methionine (SAM) is an important molecule in the cellular metabolism of mammals. In this study, we examined several of the physiological characteristics of a SAM-accumulating strain of the yeast Scheffersomyces stipitis (M12), including SAM production, ergosterol content, and ethanol tolerance. S. stipitis M12 accumulated up to 52.48 mg SAM/g dry cell weight. Proteome analyses showed that the disruption of C-24 methylation in ergosterol biosynthesis, a step mediated by C-24 sterol methyltransferase (Erg6p), results in greater SAM accumulation by S. stipitis M12 compared to the wild-type strain. A comparative proteome-wide analysis identified 25 proteins that were differentially expressed by S. stipitis M12. These proteins are involved in ribosome biogenesis, translation, the stress response, ubiquitin-dependent catabolic processes, the cell cycle, ethanol tolerance, posttranslational modification, peroxisomal membrane stability, epigenetic regulation, the actin cytoskeleton and cell morphology, iron and copper homeostasis, cell signaling, and energy metabolism.

Comparison of Proteomic, Metabolic, and Growth Profiles for Brettanomyces bruxellensis Isolates from Croatian Wines

Brettanomyces bruxellensis is one of the most important spoilage yeasts in red wine production. The aim of this paper was to investigate the diversity of B. bruxellensis isolates in regard to their proteomic, growth, and metabolic profiles. Ten isolates were obtained from several wineries in Croatian winegrowing regions during different phases of wine production. Proteomic analysis revealed 12 proteins that were expressed by all tested isolates and the reference strain. These proteins could be used as biomarkers in Dekkera/B. bruxellensis yeast identification. Five of these proteins were involved in carbohydrate metabolism (enolase, hxk2p, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, and pyruvate decarboxylase), and four were involved in protein biosynthesis (elongation factor 1-alpha, eukaryotic translation initiation factor 5a, 60s ribosomal protein 13, and putative cytosolic ribosomal protein s24). One each was involved in cellular stress responses to glucose starvation (heat shock protein ssb1); ubiquitin conjugation pathways such as transcription, proteolysis trafficking, and kinase activation (ubiquitin); or nitrogen metabolism (peptidyl-prolyl cis-trans isomerase). Isolates identified as B. bruxellensis and taken during malolactic fermentation expressed 18 similar proteins, and isolates from aging in steel vessels/wood barrels or bottled wine expressed 23 and 24 similar proteins, respectively. Growth and metabolic profiles of these isolates were evaluated in two growth media (glucose complex and wine-mimicking media). The growth profiles of the tested isolates and conversion of hydroxycinnamic acids varied among the investigated media in that the use of glucose complex medium resulted in faster growth and consumption of hydroxycinnamic acids and in higher production of volatile phenols and esters. The results obtained suggest the possibility of applying proteomic fingerprinting for the identification and differentiation of B. bruxellensis wine isolates and reveal different spoilage capacities, such as growth and metabolic profiles, of the tested isolates.