Fermentation and oxygen transfer characteristics in recombinant human growth hormone production by <i>Pichia pastoris</i> in sorbitol batch and methanol fed‐batch operation

Çalı́k, Pınar; İnankur, Bahar; Soyaslan, Elif Ş.; Şahin, Merve; Taşpınar, Hatice; Acik, Eda; Bayraktar, Eda

doi:10.1002/jctb.2292

Cited by 17 publications

(16 citation statements)

References 29 publications

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“…This is also evidenced by the increased oxygen requirements (Jungo et al, ) (Fig. C), while it has been described previously that the protein yield on methanol increases significantly when a co‐substrate is used (Çalik et al, ; Eskitoros and Çalık, ; Jungo et al, ).…”

Section: Resultssupporting

confidence: 52%

Genome‐scale metabolic model of Pichia pastoris with native and humanized glycosylation of recombinant proteins

Irani

Kerkhoven

Shojaosadati

et al. 2015

Biotech & Bioengineering

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Pichia pastoris is used for commercial production of human therapeutic proteins, and genome-scale models of P. pastoris metabolism have been generated in the past to study the metabolism and associated protein production by this yeast. A major challenge with clinical usage of recombinant proteins produced by P. pastoris is the difference in N-glycosylation of proteins produced by humans and this yeast. However, through metabolic engineering, a P. pastoris strain capable of producing humanized N-glycosylated proteins was constructed. The current genome-scale models of P. pastoris do not address native nor humanized N-glycosylation, and we therefore developed ihGlycopastoris, an extension to the iLC915 model with both native and humanized N-glycosylation for recombinant protein production, but also an estimation of N-glycosylation of P. pastoris native proteins. This new model gives a better prediction of protein yield, demonstrates the effect of the different types of N-glycosylation of protein yield, and can be used to predict potential targets for strain improvement. The model represents a step towards a more complete description of protein production in P. pastoris, which is required for using these models to understand and optimize protein production processes.

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Section: Resultssupporting

confidence: 52%

Genome‐scale metabolic model of Pichia pastoris with native and humanized glycosylation of recombinant proteins

Irani

Kerkhoven

Shojaosadati

et al. 2015

Biotech & Bioengineering

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“…A 3.0 L pilot‐scale bioreactor (BBraun, CT2‐2, Melsungen, Germany), having a working volume of 1.0 L, and consisting of temperature, pH, foam, stirring rate, feed inlet rate and dissolved oxygen controls was used. Production was carried out at N = 900 rpm, T = 30 °C 15, 16. Dissolved oxygen (DO) concentration was maintained above 20% air saturation, by enriching the inlet air with pure oxygen passing through a mass flow controller when needed 15, 16.…”

Section: Methodsmentioning

confidence: 99%

“…Production was carried out at N = 900 rpm, T = 30 °C 15, 16. Dissolved oxygen (DO) concentration was maintained above 20% air saturation, by enriching the inlet air with pure oxygen passing through a mass flow controller when needed 15, 16. To keep pH constant throughout the bioprocess, 25% ammonia solution, NH 4 OH, was used 15, 16…”

Section: Methodsmentioning

confidence: 99%

Influence of pH on recombinant human growth hormone production by Pichia pastoris

Çalı́k

Bayraktar

İnankur

et al. 2010

J of Chemical Tech & Biotech

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“…compared the rhGH production potentials of Mut S and Mut + strains in batch experiments and reported 0.16 g L −1 rhGH production by Mut + on complex medium, and 0.11 g L −1 rhGH production by Mut S in medium containing C Gly = 30 g L −1 and C M = 1% (v/v). Çalık et al . investigated the effects of different methanol feeding rates and pH conditions on rhGH production by P. pastoris hGH ‐Mut + strain, and obtained the highest AOX 1 and hGH expression levels and rhGH concentration ( C rhGH = 0.27 g L −1 ) at controlled pH = 5.0.…”

Section: Introductionmentioning

confidence: 99%

Methanol feeding strategy design enhances recombinant human growth hormone production by Pichia pastoris

Güneş

Boy

Ata

et al. 2015

J of Chemical Tech & Biotech

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BACKGROUND The effects of designed methanol and co‐substrate feeding strategies on fed‐batch recombinant human growth hormone (rhGH) production by Pichia pastoris hGH‐Mut+ were investigated. RESULTS The highest cell concentration was obtained as 157 g L−1 with the exponential feeding strategy where methanol was fed at a pre‐calculated feeding rate based on pre‐determined µ0 = 0.03 h−1, and mannitol concentration was kept constant at 50 g L−1 for t=0–6 h. The highest rhGH concentration was obtained as 1.3 g L−1 at t=54 h with the methanol‐stat feeding strategy, where no co‐substrate was used and the methanol concentration was kept at 5 g L−1 in the medium during the production phase. Thereafter, in another methanol‐stat feeding strategy with pulse‐fed sorbitol, the highest rhGH concentration was obtained as 1.2 g L−1 at t=42 h. CONCLUSION The methanol‐stat fed‐batch method keeping methanol concentration constant at 5 g L−1 in the production medium increased rhGH production at least 2‐fold compared with that of the exponential feeding strategies. In the designed methanol‐stat feeding strategies, use of sorbitol as co‐substrate decreased methanol demand of the cells 2‐fold, increased the overall product yield on the substrate 1.6‐fold, shortened the cultivation time by 12 h and increased the productivity. Thus, methanol‐stat feeding with co‐substrate sorbitol was found to be a favorable fed‐batch strategy for rhGH production by Pichia pastoris under alcohol oxidase I promoter. © 2014 Society of Chemical Industry

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Fermentation and oxygen transfer characteristics in recombinant human growth hormone production by Pichia pastoris in sorbitol batch and methanol fed‐batch operation

Cited by 17 publications

References 29 publications

Genome‐scale metabolic model of Pichia pastoris with native and humanized glycosylation of recombinant proteins

Genome‐scale metabolic model of Pichia pastoris with native and humanized glycosylation of recombinant proteins

Influence of pH on recombinant human growth hormone production by Pichia pastoris

Methanol feeding strategy design enhances recombinant human growth hormone production by Pichia pastoris

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