STUDIES ON PLASMID STABILITY, CELL-METABOLISM AND SUPEROXIDE-DISMUTASE PRODUCTION BY PGK- STRAINS OF SACCHAROMYCES-CEREVISIAE

A double mutant sod1/pgk1 strain of Saccharomyces cerevisiae has been constructed in order to investigate the effects of different environmental conditions on yeast physiology, plasmid stability, and superoxide dismutase (SOD) production. Strains were transformed with yeast episomal plasmids (YEp) containing both PGK1 and SOD1 genes and were grown on fermentable carbon sources and under vigorous aeration. Under these conditions, the presence of the PGK1 gene was made essential for growth and both genes were efficiently expressed. However, plasmid-borne PGK1 was found not to increase the stability of YEp vectors in batch cultures of Pgk- cells. Paradoxically, plasmid stability increased during the respiratory phase of growth. An investigation of the metabolism of Pgk- cells demonstrated that these glycolytic pathway mutants do not appreciably metabolize glycerol. Thus Pgk+, plasmid-containing, cells have a selective advantage during the respiratory phase of batch growth since they can utilize both glycerol and ethanol.