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

Z. A. Irani ; Eduard Kerkhoven (Institutionen för biologi och bioteknik, Systembiologi) ; S. A. Shojaosadati ; Jens B. Nielsen (Institutionen för biologi och bioteknik, Systembiologi)
Biotechnology and Bioengineering (0006-3592). Vol. 113 (2016), 5, p. 961-969.
[Artikel, refereegranskad vetenskaplig]

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.

Nyckelord: genome scale metabolic model, humanized glycosylation, Pichia pastoris, recombinant protein production

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Denna post skapades 2016-05-02. Senast ändrad 2017-01-17.
CPL Pubid: 235744


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Institutionen för biologi och bioteknik, Systembiologi



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