Mutant and deletion strains of the yeast Saccharomyces cerevisiae having one/several auxotrophies are largely used in the development of recombinant strains for heterologous protein production because they ensure maintenance of plasmids with selectable markers. The production is usually carried out by culturing the recombinant strain in aerated fed-batch, where sugar limitation achieves high yields of biomass and product. In a previous work, it was evidenced that growth of the auxotrophic S. cerevisiae BY4741 (MATa, ura30, leu20, met150, his31) engineered for human IL-1 production, and employed in aerated fed-batch, early arrested even in the presence of a correct nutritional complementation (being specific nutrients for genetically uncomplemented auxotrophies provided in no growthlimiting amounts). It was assumed that this behaviour may depend on the high number of auxotrophies, since the prototrophic S288C, from which BY4741 derives, showed a typical performance under the same cultivation mode. Therefore, a systematic investigation on the effect of auxotrophies on yeast growth in aerated fed-batch was carried out. Four isogenic strains of the CEN.PK family, with a progressively increasing number of auxotrophies (from one to four) were assayed under fed-batch conditions and a proper nutritional complementation. Feeding to the reactor was exponentially increased imposing a specific growth rate below the critical one. The behaviour of the auxotrophic strains was compared with that of the isogenic prototrophic strain. by evaluating the capacity to keep the specific growth rate chosen. A clear correlation among optimum growth and number of auxotrophies has been found. Furthermore we have investigated the possible effect of the type of auxotrophy (ura- or leu-) on the strain performance, monitoring as well cell viability of each strain. The study is a contribution to know the phenotypic effects of auxotrophies in yeast and can have implications for biotechnological applications.

Effect of auxotrophies on yeast growth in aerated fed-batch reactor

PACIELLO, LUCIA;LANDI, CARMINE;PARASCANDOLA, Palma
2010

Abstract

Mutant and deletion strains of the yeast Saccharomyces cerevisiae having one/several auxotrophies are largely used in the development of recombinant strains for heterologous protein production because they ensure maintenance of plasmids with selectable markers. The production is usually carried out by culturing the recombinant strain in aerated fed-batch, where sugar limitation achieves high yields of biomass and product. In a previous work, it was evidenced that growth of the auxotrophic S. cerevisiae BY4741 (MATa, ura30, leu20, met150, his31) engineered for human IL-1 production, and employed in aerated fed-batch, early arrested even in the presence of a correct nutritional complementation (being specific nutrients for genetically uncomplemented auxotrophies provided in no growthlimiting amounts). It was assumed that this behaviour may depend on the high number of auxotrophies, since the prototrophic S288C, from which BY4741 derives, showed a typical performance under the same cultivation mode. Therefore, a systematic investigation on the effect of auxotrophies on yeast growth in aerated fed-batch was carried out. Four isogenic strains of the CEN.PK family, with a progressively increasing number of auxotrophies (from one to four) were assayed under fed-batch conditions and a proper nutritional complementation. Feeding to the reactor was exponentially increased imposing a specific growth rate below the critical one. The behaviour of the auxotrophic strains was compared with that of the isogenic prototrophic strain. by evaluating the capacity to keep the specific growth rate chosen. A clear correlation among optimum growth and number of auxotrophies has been found. Furthermore we have investigated the possible effect of the type of auxotrophy (ura- or leu-) on the strain performance, monitoring as well cell viability of each strain. The study is a contribution to know the phenotypic effects of auxotrophies in yeast and can have implications for biotechnological applications.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11386/3808879
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