title: Genome-scale metabolic model of Pichia pastoris with native and humanized glycosylation of recombinant proteins.
abstract: 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.
authors.affiliation: Biotechnology Group, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, Iran.
authors.forename: Zahra Azimzadeh
authors.initials: ZA
authors.lastname: Irani
authors.affiliation: Systems and Synthetic Biology, Department of Biology and Biological Engineering, Chalmers University of Technology, Göteborg, Sweden.
authors.forename: Eduard J
authors.initials: EJ
authors.lastname: Kerkhoven
authors.affiliation: Biotechnology Group, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, Iran. shoja_sa@modares.ac.ir.
authors.forename: Seyed Abbas
authors.initials: SA
authors.lastname: Shojaosadati
authors.affiliation: Systems and Synthetic Biology, Department of Biology and Biological Engineering, Chalmers University of Technology, Göteborg, Sweden. nielsenj@chalmers.se.|Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark. nielsenj@chalmers.se.
authors.forename: Jens
authors.initials: J
authors.lastname: Nielsen
mesh_terms: D006031:Glycosylation; D006801:Humans; D007218:Industrial Microbiology / Q000379:methods; D060847:Metabolic Engineering / Q000379:methods; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins
keywords: Pichia pastoris; genome scale metabolic model; humanized glycosylation; recombinant protein production