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Information × Registration Number 0216U002463, 0111U002630 , R & D reports Title Mechanisms of regulation of autophagy, stress response and synthesis of biologically active compounds in yeasts. popup.stage_title Head Sibirny Andriy Andrijovych, Registration Date 17-03-2016 Organization Institute of Cell Biology of National Academy of Sciences of Ukraine popup.description2 Potential protein partners of protein Atg 28 which is involved in the majority of autophagic pathways in yeast Pichia pastoris, particularly protein Atg17, which regulates activity of Atg1 and is necessary for the formation of a full-size autophagosome and a novel protein, Atg35, involved in the formation of specific MIPA-structures during micropexophagy were identified. New genes, MON1, CCZ1 and YPT7 of methylotrophic yeast P. pastoris, have been identified that are involved in pexophagy, in the pathway of transportation of hydrolases from the cytosol to the vacuole (Svt-pathway) and in nonspecific macroautophagy. Also, GSS1 gene of this yeast species encoding the glucose sensor was identified. Its deletion also leads towards pexophagy. The mechanisms of response to stress caused by alcohols in methylotrophic yeast Hansenula polymorpha were investigated. A new gene ETT1 has been identified, and its deletion leads to hypersensitivity to ethanol, while overexpression, on the contrary, to the increased resistance to ethanol and elevated temperatures. The overexpression of FPS1 gene induced the increased resistance of this yeast species to ethanol and higher alcohols, in particular, to isobutanol. H. polymorpha mutants with low and high content of the tripeptide glutathione in cells were selected, which can be used for further breeding of overproducers of this tripeptide. Strains with increased synthesis of ethanol from pentose xylose were constructed. This was achieved by overexpression of genes of first three initial reactions of xylose catabolism, XYL1m, XYL2 and XYL3, as well as by deletion or disruption of the gene ATG13 involved in the autophagy initiation. In order to study the possibility of hydrogen production in bacteria we used recombinant clones of chemolitotrophic bacterium Shewanella oneidensis with deleted genes hydA and hyaB encoding endogenous hydrogenases of this species which produce labeled hydrogenase of Pseudotrichonympha grassii. It was found that the synthesis of riboflavin in the yeast Candida famata can be increased by modification and subsequent overexpression of PRS3 and ADE4 genes (encoding phosphoribosil pyrophosphate synthetase and phosphoribosil pyrophosphate amidotransferase, respectively) that are involved in GTP biosynthesis de novo. It was shown that the increasing of riboflavin overproduction can be achieved through overexpression of a SEF1 gene identified previously, which encodes transcription activator. qRT-PCR showed that SEF1 in C. famata and P. stipitis is regulated by iron deprivation in the medium. The potential binding sites for protein Sef1 in the promoter of RIB1 gene were identified using yeast monohybrid system. For the first time, we constructed recombinant strains of C. famata, which are able to overproduce a flavin nucleotide FAD without the addition of costly exogenous precursors. This was achieved by means of overexpression of FAD1 gene, encoding FAD synthetase, in the previously constructed strain that is able to overproduce other flavin nucleotide, FMN. Product Description popup.authors Борецький Ю.Р. Булботка Н. Гринів О.Б. Дмитрук К.В. Дмитрук О.В. Куриленко О.О. Кшановська Б.В. Левків К.О. Лизак О.О. Лужецький Т.Б. Муращенко Л.Р. Пиняга Ю.В. Рачкевич Н.О. Семків М.В. Фаюра Л.Р. Федоренко В.О. Федорович Д.В. Юрків М. Яковенко Г.Й. Яцишин В.Ю. popup.nrat_date 2020-04-02 Close