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

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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-1β (IL-1β), aiming at obtaining high cell densities and optimizing IL-1β 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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Abbreviations

\(\beta\) :

Inhibition factor of glucose on ethanol consumption

\(E\) :

Ethanol concentration at time \(t\) (g L−1)

\(E_{0}\) :

Ethanol concentration at the end of batch phase/beginning of fed-batch phase (g L−1)

\(F\) :

Glucose flow rate at time \(t\) (L h−1)

\(F_{0}\) :

Glucose flow rate at the beginning of fed-batch phase (L h−1)

\(G\) :

Residual glucose concentration at time \(t\) (g L−1)

\(G_{0}\) :

Residual glucose concentration at the beginning of fed-batch phase (g L−1)

\(G_{\text{R}}\) :

Glucose concentration in the feeding (g L−1)

\(k_{\text{d}}\) :

Specific death rate (h−1)

\(\left( {k_{S} } \right)_{E}\) :

Ethanol saturation constant (g L−1)

\(\left( {\mu_{ \hbox{max} } } \right)_{E}\) :

Maximum specific growth rate on ethanol (h−1)

\(\left( {\mu_{G} } \right)_{i}\) :

Specific growth rate on glucose in the i th interval (h−1)

\(t\) :

Arbitrary time of fed-batch run (h−1)

\(t_{\text{EC}}\) :

Time of fed-batch run when ethanol finished to be consumed and the variation of specific growth rate on glucose equation occurs (h)

\(t_{\text{EP}}\) :

Time of fed-batch run when ethanol begins to be accumulated (h)

\(V\) :

Broth culture volume (L)

\(X\) :

Viable biomass concentration at time \(t\) (g cell d.w. L−1)

\(X_{0}\) :

Initial viable biomass concentration (g cell d.w. L−1)

\(\left( {Y_{X/G} } \right)_{i}\) :

Biomass yield on glucose in the i-th interval of fed-batch run (g cell d.w. g glucose−1)

\(\left( {Y_{X/G} } \right)_{1}\) :

Biomass yield on glucose in the interval 0 ≤ t < t EC (g cell d.w. g glucose−1)

\(\left( {Y_{X/G} } \right)_{2}\) :

Biomass yield on glucose in the interval t ≥ t EC (g cell d.w. g glucose−1)

\(\left( {Y_{X/E} } \right)_{S}\) :

Biomass yield on ethanol as substrate in the interval 0 ≤ t < t EC (g cell d.w. g ethanol−1)

\(\left( {Y_{E/X} } \right)_{P}\) :

Ethanol produced per unit of biomass in the interval t ≥ t EP (g ethanol g cell d.w.−1)

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Acknowledgments

This research was supported by the University of Salerno funds (FARB, 2011) to Palma Parascandola in the framework of the research project “Ottimizzazione e modellazione di sistemi HCDC (high cell-density cultivation) per la produzione di masse microbiche da lievito: parametri biologici e condizioni di processo”.

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Authors and Affiliations

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

    Carmine Landi, Lucia Paciello & Palma Parascandola

  2. Dep. Biologia, Università degli Studi di Napoli “Federico II”, Via Cinthia, 80100, Naples, Italy

    Elisabetta de Alteriis

  3. Dep. Biotecnologie e Bioscienze, Università Milano-Bicocca, Piazza Della Scienza, 20126, Milan, Italy

    Luca Brambilla

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  1. Carmine Landi
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  2. Lucia Paciello
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  3. Elisabetta de Alteriis
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  4. Luca Brambilla
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  5. Palma Parascandola
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Correspondence to Palma Parascandola.

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Landi, C., Paciello, L., de Alteriis, E. et al. High cell density culture with S. cerevisiae CEN.PK113-5D for IL-1β production: optimization, modeling, and physiological aspects. Bioprocess Biosyst Eng 38, 251–261 (2015). https://doi.org/10.1007/s00449-014-1264-8

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