| Title: | [Enhancement of Coprinus cinereus peroxidase in Pichia pastoris by co-expression chaperone PDI and Ero1] |
| Author(s): | Chen F; Hu M; Jiang X; Tao Y; Huang J; |
| Journal Title: | Sheng Wu Gong Cheng Xue Bao |
| ISSN/ISBN: | 1000-3061 (Print) 1000-3061 (Linking) |
| 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" |
| Keywords: | Coprinus/*enzymology Culture Media Cytoplasm Fermentation Glycoproteins/*metabolism Mass Spectrometry Mating Factor Oxidoreductases Acting on Sulfur Group Donors/*metabolism Peptides Peroxidases/*biosynthesis Pichia/*metabolism Protein Disulfide-Isomerase; |
| Notes: | "MedlineChen, Fei Hu, Meirong Jiang, Xianzhang Tao, Yong Huang, Jianzhong chi Research Support, Non-U.S. Gov't China 2016/04/21 Sheng Wu Gong Cheng Xue Bao. 2015 Dec; 31(12):1682-9" |
