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Enhancement of Coprinus cinereus peroxidase in Pichia pastoris by co-expression chaperone PDI and Ero1 / 生物工程学报
Chinese Journal of Biotechnology ; (12): 1682-1689, 2015.
Article in Zh | WPRIM | ID: wpr-337467
Responsible library: WPRO
ABSTRACT
The 1,095 bp gene encoding peroxidase from Coprinus cinereus was synthesized and integrated into the genome of Pichia pastoris with a highly inducible alcohol oxidase. The recombinant CiP (rCiP) fused with the a-mating factor per-pro leader sequence derived from Saccharomyces cerevisiae was secreted into the culture medium and identified as the target protein by mass spectrometry, confirming that a C. cinereus peroxidase (CiP) was successfully expressed in P. pastoris. The endoplasmic reticulum oxidoreductase 1 (Ero1) and protein disulfide isomerase (PDI) were co-expressed with rCiP separately and simultaneously. Compared with the wild type, overexpression of PDI and Erol-PDI increaseed Cip activity in 2.43 and 2.6 fold and their activity reached 316 U/mL and 340 U/mL respectively. The strains co-expressed with Erol-PDI was used to high density fermentation, and their activity reached 3,379 U/mL, which was higher than previously reported of 1,200 U/mL.
Subject(s)
Full text: 1 Index: WPRIM Main subject: Peptides / Peroxidases / Pichia / Saccharomyces cerevisiae / Mass Spectrometry / Glycoproteins / Protein Folding / Coprinus / Protein Disulfide-Isomerases / Culture Media Type of study: Prognostic_studies Language: Zh Journal: Chinese Journal of Biotechnology Year: 2015 Type: Article
Full text: 1 Index: WPRIM Main subject: Peptides / Peroxidases / Pichia / Saccharomyces cerevisiae / Mass Spectrometry / Glycoproteins / Protein Folding / Coprinus / Protein Disulfide-Isomerases / Culture Media Type of study: Prognostic_studies Language: Zh Journal: Chinese Journal of Biotechnology Year: 2015 Type: Article