A Model for β‐Galactosidase Production with a Recombinant Yeast <i>Saccharomyces cerevisiae</i> in Fed‐Batch Culture

Hardjito, Linawati; Greenfield, P. F.; Lee, Peter L.

doi:10.1021/bp00016a006

Cited by 12 publications

(6 citation statements)

References 41 publications

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“…With increasing understanding of these factors, it is possible to enhance plasmid stability by manipulating plasmid composition and structure [23], changing the genetic and physiological properties of host cells [2], and manipulating the environmental conditions [17,24,25], dilution rate [13,17,29] and bioreactor operation modes [13,15].…”

Section: Introductionmentioning

confidence: 99%

Plasmid stability and kinetics of continuous production of glucoamylase by recombinant Saccharomyces cerevisiae in an airlift bioreactor

Kilonzo

Margaritis

Bergougnou

2009

J Ind Microbiol Biotechnol

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Production of glucoamylase by recombinant Saccharomyces cerevisiae C468/pGAC9 (ATCC 20690) in a continuous stirred tank bioreactor was studied at different dilution rates. Plasmid stability was found to be growth (dilution rate) dependent; it increased with the dilution rate. Bioreactor productivity and specific productivity also increased with the dilution rate. A kinetic equation was used to model the plasmid stability kinetics. The growth rate ratio between plasmid-carrying and plasmid-free cells decreased from 1.397 to 1.215, and segregational instability or probability of plasmid loss from each cell division decreased from 0.059 to 0.020 as the dilution rate increased from 0.10 to 0.37 1/h. The specific growth rates increased with dilution rate, while the growth rate difference between plasmid-carrying and plasmid-free cell populations was negligible. This was attributed to the low copy number of the hybrid plasmid pGAC9. Thus, the growth rate had no significant effect on plasmid instability. The proposed kinetics was consistent with experimental results, and the model simulated the experimental data well.

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Section: Introductionmentioning

confidence: 99%

Plasmid stability and kinetics of continuous production of glucoamylase by recombinant Saccharomyces cerevisiae in an airlift bioreactor

Kilonzo

Margaritis

Bergougnou

2009

J Ind Microbiol Biotechnol

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“…Thus, they were unable to simulate the dynamics of cell growth, substrate consumption and gene product formation. In recent years, relatively few studies on modelling of recombinant yeast addressed all of these aspects [5,6]. By utilizing the available knowledge on the behaviour of recombinant S. cerevisiae, Coppella and Dhurjati [5] formulated a detailed structured model for describing cell growth, plasmid segregation and heterogeneous protein production.…”

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confidence: 99%

“…Although this model successfully described the observed results for the fermentation of recombinant S. cerevisiae strain AB103.2/pYaEGF-25 in both batch and fed-batch bioreactors, its complexity poses some practical dif®culties for engineering applications. An unstructured model was developed to describe the growth and product formation of a recombinant S. cerevisiae expressing b-galactisidase in fed-batch bioreactor [6]. This model was based on Monod equation and required 10 equations and 12 parameters.…”

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confidence: 99%

Mathematical model for aerobic culture of a recombinant yeast

Zhang

Scharer

Moo‐Young

1997

Bioprocess Engineering

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A mathematical model was formulated to simulate cell growth, plasmid loss and recombinant protein production during the aerobic culture of a recombinant yeast S. cerevisiae. Model development was based on three simpli®ed metabolic events in the yeast: glucose fermentation, glucose oxidation and ethanol oxidation. Cell growth was expressed as a composite of these metabolic events. Their contributions to the total speci®c growth rate depended on the activities of the pacemaker enzyme pools of the individual pathways. The pacemaker enzyme pools were regulated by the speci®c glucose uptake rate. The effect of substrate concentrations on the speci®c growth rate was described by a modi®ed Monod equation. It was assumed that recombinant protein formation is only associated with oxidative pathways. Plasmid loss kinetics was formulated based on segregational instability during cell division by assuming constant probability of plasmid loss. Experiments on batch fermentation of recombinant S. cerevisiae C468/pGAC9 (ATCC 20690), which expresses Aspergillus awamori glucoamylase gene and secretes glucoamylase into the extracellular medium, were carried out in an airlift bioreactor in order to evaluate the proposed model. The model successfully predicted the dynamics of cell growth, glucose consumption, ethanol metabolism, glucoamylase production and plasmid instability. Excellent agreement between model simulations and our experimental data was achieved. Using published experimental data, model agreement was also found for other recombinant yeast strains. In general, the proposed model appears to be useful for the design, scale-up, control and optimization of recombinant yeast bioprocesses.

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“…This model was developed from a model of Hardjito et al [10] with some modi®cations in the glucose metabolism.…”

Section: Methodsmentioning

confidence: 99%

Discontinuities in continuous cultures studied with a recombinant Saccharomyces cerevisiae and two computer models

Ryszczuk

Grøn

Carlsen

et al. 1997

Bioprocess Engineering

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Chemostat cultures and other continuous cultures, where the feed to the bioreactor was divided in pulses while maintaining the overall dilutions rate by the increased¯ow in the pulses, were studied by cultivations with Saccharomyces cerevisiae JG176 and by simulations with two computer models, Yeast model SG176 and Yeast model MC176. All three systems gave interesting response surfaces. Deviations from an ideal chemostat may have signi®cant effects on volumetric productivity, which for production of the recombinant protein, proteinase A by Saccharomyces cerevisiae JG176 was positive, while the productivity of biomass and ethanol decreased. In simulations with both models pulsing caused lower production of biomass and ethanol. In simulations with one of the models the effects of pulsing on productivity of a protein were also negative, whereas simulations with the other model suggested clear positive effects of pulsing on a production of a protein though with a somewhat different response surface than with the experiments with Saccharomyces cerevisiae JG176.

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A Model for β‐Galactosidase Production with a Recombinant Yeast Saccharomyces cerevisiae in Fed‐Batch Culture

Cited by 12 publications

References 41 publications

Plasmid stability and kinetics of continuous production of glucoamylase by recombinant Saccharomyces cerevisiae in an airlift bioreactor

Plasmid stability and kinetics of continuous production of glucoamylase by recombinant Saccharomyces cerevisiae in an airlift bioreactor

Mathematical model for aerobic culture of a recombinant yeast

Discontinuities in continuous cultures studied with a recombinant Saccharomyces cerevisiae and two computer models

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