A structured kinetic model for recombinant protein production by Mut(+) strain of Pichia pastoris

Date

2009-12-01

Author

Celik, Eda
Çalık, Pınar
Oliver, Stephen G.

Metadata

Show full item record

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Item Usage Stats

101
views

0
downloads

A structured kinetic biochemical model for Pichia pastoris growth and recombinant protein production, able to predict r-protein production and optimize the feeding strategy is reported. Pichia pastoris was divided into three compartments, the recombinant product human erythropoietin (rHuEPO) was considered in one compartment, alcohol oxidase and extracellular protease enzymes were assembled in the enzymatic compartment, and rest of the cell components (including intracellular proteases) were gathered in the cellular compartment. After parameter identification by the simplex method, sensitivity analysis was carried out to confirm that the global optimum values were reached for the parameters of the model. The model successfully simulated the dynamic changes and the resulting rHuEPO production caused by variations in methanol feeding rate, even in the lag and the stationary phases. The model successfully predicted the optimal feeding strategies and has great potential for use in process simulation and control of glycosylated recombinant protein production by Pichia pastoris cells.

Subject Keywords

Pichia pastoris, Fed-batch fermentation, Bioprocessing, Biochemical engineering, Parameter identification, Mathematical modelling

URI

https://hdl.handle.net/11511/41730

Journal

CHEMICAL ENGINEERING SCIENCE

DOI

https://doi.org/10.1016/j.ces.2009.08.009

Collections

Department of Chemical Engineering, Article

Suggestions

OpenMETU
Core

Defined and complex medium based feeding strategy development for recombinant human growth hormone production by P. pastoris under GAP promoter Hoxha, Bebeta; Çalık, Pınar; Özdamar, Tunçer H.; Department of Chemical Engineering (2016) The objective of this study is to investigate different feeding strategies leading to higher recombinant human growth hormone (rhGH) production by Pichia pastoris, driven under constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene. rhGH production took place in a pilot bioreactor where glucose and molasses were examined as carbon sources. For batch phase, which is the first production phase of the reactor, all experiments share the same characteristics where glycerol was utilized as the...
Double promoter expression systems for recombinant protein production by industrial microorganisms Ozturk, Sibel; Ergun, Burcu Gunduz; Çalık, Pınar (2017-10-01) Using double promoter expression systems is a promising approach to increase heterologous protein production. In this review, current double promoter expression systems for the production of recombinant proteins (r-proteins) by industrially important bacteria, Bacillus subtilis and Escherichia coli; and yeasts, Saccharomyces cerevisiae and Pichia pastoris, are discussed by assessing their potentials and drawbacks. Double promoter expression systems need to be designed to maintain a higher specific product f...
Identification of Novel Reference Genes Based on MeSH Categories Ersahin, Tulin; ÇARKACIOĞLU, Levent; Can, Tolga; Konu, Ozlen; Atalay, Mehmet Volkan; Atalay, Rengül (2014-03-28) Transcriptome experiments are performed to assess protein abundance through mRNA expression analysis. Expression levels of genes vary depending on the experimental conditions and the cell response. Transcriptome data must be diverse and yet comparable in reference to stably expressed genes, even if they are generated from different experiments on the same biological context from various laboratories. In this study, expression patterns of 9090 microarray samples grouped into 381 NCBI-GEO datasets were invest...
In-silico determination of Pichia pastoris signal peptides for extracellular recombinant protein production Massahi, Aslan; Çalık, Pınar (2015-01-07) In-silico identified novel secretory signal peptides (SPs) are required in vivo to achieve efficient transfer or to prevent other cellular proteins from interfering with the process in extracellular recombinant protein (r-protein) production. 56 endogenous and exogenous secretory SPs, have been used or having the potential to be used in Pichia pastoris for r-protein secretion, were analyzed in-silico using the softwares namely SignalP4.1, Phobius, WolfPsort0.2, ProP1.0, and NetNGlyc1.0. Among the predicted ...
A multi-layered graphical model of the relation among SNPS, GENES, and pathways based on subgraph search Ersoy, Gökhan; Aydın Son, Yeşim; Can, Tolga; Department of Bioinformatics (2015) The analysis of Single Nucleotide Polymorphisms (SNPs) through Genome Wide Association Studies (GWAS) presents great potential for describing disease loci and gaining insight into the underlying etiology of diseases. Recently described combined p-value approach allows identification of associations at gene and pathway level. The integrated programs like METU-SNP produce simple lists of either SNP id/gene id/pathway title and their p-values and significance status or SNP id/disease id/pathway information. In...

Citation Formats

E. Celik, P. Çalık, and S. G. Oliver, “A structured kinetic model for recombinant protein production by Mut(+) strain of Pichia pastoris,” CHEMICAL ENGINEERING SCIENCE, pp. 5028–5035, 2009, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/41730.