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High cell density culture with S. cerevisiae CEN.PK113-5D for IL-1{Beta} production: optimization, modeling, and physiological aspects.

Landi C, Paciello L, de Alteriis E, Brambilla L, Parascandola P.

Bioprocess Biosyst Eng. 2015 ;38(2):251-61.

Dep. Ingegneria Industriale, Università Di Salerno, Via Giovanni Paolo II, 132 Fisciano, 84084, Salerno, Italy.

 

Abstract

Saccharomyces cerevisiae CEN.PK113-5D, a strain auxotrophic for uracil belonging to the CEN.PK family of the yeast S. cerevisiae, was cultured in aerated fed-batch reactor as such and once transformed to express human interleukin-1{Beta} (IL-1{Beta}), aiming at obtaining high cell densities and optimizing IL-1{Beta} production. Three different exponentially increasing glucose feeding profiles were tested, all of them “in theory” promoting respiratory metabolism to obtain high biomass/product yield. A non-structured non-segregated model was developed to describe the performance of S. cerevisiae CEN.PK113-5D during the fed-batch process and, in particular, its capability to metabolize simultaneously glucose and ethanol which derived from the precedent batch growth. Our study showed that the proliferative capacity of the yeast population declined along the fed-batch run, as shown by the exponentially decreasing specific growth rates on glucose. Further, a shift towards fermentative metabolism occurred. This shift took place earlier the higher was the feed rate and was more pronounced in the case of the recombinant strain. Determination of some physiological markers (acetate production, intracellular ROS accumulation, catalase activity and cell viability) showed that neither poor oxygenation nor oxidative stress was responsible for the decreased specific growth rate, nor for the shift to fermentative metabolism.

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Saccharomyces_cerevisiae