Semkiv M. Metabolic engineering of yeast Saccharomyces cerevisiae for construction of the efficient fuel ethanol and glycerol producers

Aim of the study: The purpose of this work was to obtain strains of baker's yeast S. cerevisiae, capable of increased production of ethanol or glycerol during alcoholic fermentation. Methods of research: The genetic, biochemical and microbiological methods of research were used in this work. As a result of the work, a vector for gene expression with a modified selection marker (kanMX4 gene) was developed that provides multicopy integration in the yeast of S. cerevisiae. On the basis of the industrial ethanol producer, a recombinant strain of S. cerevisiae with an overexpression of the PHO8 gene (coding for alkaline phosphatase) has been constructed, which is characterized by a 6% increase in ethanol production. Expression of apy E. coli gene encoding apyrase, as well as activation of futile cycles, have been found to have a positive effect on ethanol production during alcoholic fermentation in S. cerevisiae. A glycerol-producing S. cerevisiae strain has been obtained, which synthesized 9 or 4.7 more glycerol than a wild type strain during alcoholic fermentation in microaerobic or anaerobic conditions, respectively.