The auxotrofic S. cerevisiae BY4741, carrying the fusion PIR4-IL1β, and able to secrete the human interleukin-1β into the culture medium, has been employed in an aerated bioreactor working as a fed-batch. Notwithstanding proper formulation of the culture medium, the performance of the host strain BY4741 [Pir4-IL1β] in fed-batch was not satisfactory: biomass density was far from that expected, glucose and ethanol accumulated during the runs. To test if the oxidative stress was responsible for the observed behaviour, the mutant BY4741 Δyca1, deleted for the YCA1 gene, coding for a caspase-like protein involved in yeast apoptosis, was transformed with the expression cassette containing the fusion PIR4-IL1β, and assayed in the same aerated fed-batch system. The different performances exhibited by both BY4741[Pir4-IL1β] and BY4741 Δyca1 [Pir4-IL1β], together with the study of death kinetics during operation runs, evidenced the relevance of strain viability in the operative conditions employed.

Heterologous protein production by yeast in aerated fed-batch cultures:relevance of the host strain viability.

PACIELLO, LUCIA;
2008-01-01

Abstract

The auxotrofic S. cerevisiae BY4741, carrying the fusion PIR4-IL1β, and able to secrete the human interleukin-1β into the culture medium, has been employed in an aerated bioreactor working as a fed-batch. Notwithstanding proper formulation of the culture medium, the performance of the host strain BY4741 [Pir4-IL1β] in fed-batch was not satisfactory: biomass density was far from that expected, glucose and ethanol accumulated during the runs. To test if the oxidative stress was responsible for the observed behaviour, the mutant BY4741 Δyca1, deleted for the YCA1 gene, coding for a caspase-like protein involved in yeast apoptosis, was transformed with the expression cassette containing the fusion PIR4-IL1β, and assayed in the same aerated fed-batch system. The different performances exhibited by both BY4741[Pir4-IL1β] and BY4741 Δyca1 [Pir4-IL1β], together with the study of death kinetics during operation runs, evidenced the relevance of strain viability in the operative conditions employed.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11386/2288962
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