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Gewählte Publikation:

Heiss, S; Maurer, M; Hahn, R; Mattanovich, D; Gasser, B; .
(2013): Identification and deletion of the major secreted protein of Pichia pastoris.
Appl Microbiol Biotechnol. 2013; 97(3):1241-1249 FullText FullText_BOKU

Abstract:
A major contaminating host cell protein was identified in fed batch cultures of Pichia pastoris producing an antibody Fab fragment. Purification and peptide sequencing identified this protein to be related to the cysteine-rich secretory protein family. The same protein was also observed as one of the most abundantly secreted proteins in chemostat cultures of a wild type P. pastoris strain. It has an apparent molecular weight of 65 kDa, 2-fold higher than predicted from the amino acid sequence, which is due to high O-glycosylation. It was denominated extracellular protein X 1 (Epx1), as no clear function could be attributed to it. The EPX1 gene is upregulated in different stress situations, and the respective deletion strain was more susceptible than the wild type to the cell wall damaging agents Calcofluor white and Congo red. The EPX1 deletion strain (Delta epx1) was evaluated for its suitability for recombinant protein production. No significant difference in growth and product formation was observed between the wild type and the Delta epx1 strain. Batch purification of a Fab fragment produced in the Delta epx1 strain highlighted its superior purity due to the decreased host cell protein load.
Autor*innen der BOKU Wien:
Gasser Brigitte
Hahn Rainer
Heiss Silvia
Mattanovich Diethard
BOKU Gendermonitor:

Find related publications in this database (using NML MeSH Indexing)
Biological Products/isolation & purification;Fungal Proteins/genetics;Fungal Proteins/secretion*;Gene Deletion*;Metabolic Engineering/methods*;Pichia/genetics;Pichia/metabolism*;Recombinant Proteins/isolation & purification;Technology, Pharmaceutical/methods*;

Find related publications in this database (Keywords)
Pichia pastoris
Extracellular protein
Host cell protein
Recombinant protein
Purification
CRISP family


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