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Enhanced Secretion of Heterologous Proteins in Pichia pastoris Following Overexpression of Saccharomyces cerevisiae Chaperone Proteins
- Source :
- Biotechnology Progress. 22:1090-1095
- Publication Year :
- 2006
- Publisher :
- Wiley, 2006.
-
Abstract
- In Pichia pastoris, secretory proteins are folded and assembled in the endoplasmic reticulum (ER). However, upon introduction of foreign proteins, heterologous proteins are often retained in the cytoplasm or in the ER as a result of suboptimal folding conditions, leading to protein aggregation. The Hsp70 and Hsp40 chaperone families in the cytoplasm or in ER importantly regulate the folding and secretion of heterologous proteins. However, it is not clear which single chaperone is most important or which combination optimally cooperates in this process. In the present study we evaluated the role of the chaperones Kar2p, Sec63, YDJ1p, Ssalp, and PDI from Saccharomyces cerevisiae. We found that the introduction of Kar2p, Ssalp, or PDI improves protein secretion 4-7 times. In addition, we found that the combination chaperones of YDJlp/ PDI, YDJlp/Sec63, and Kar2p/PDI synergistically increase secretion levels 8.7, 7.6, and 6.5 times, respectively. Therefore, additional integration of chaperone genes can improve the secretory expression of the heterologous protein. Western blot experiments revealed that the chaperones partly relieved the secretion bottleneck resulting from foreign protein introduction in P. pastoris. Therefore, the findings from the present study demonstrate the presence of a network of chaperones in vivo, which may act synergistically to increase recombinant protein yields.
- Subjects :
- Saccharomyces cerevisiae Proteins
biology
Recombinant Fusion Proteins
Endoplasmic reticulum
Heterologous
Saccharomyces cerevisiae
Protein aggregation
Protein Engineering
biology.organism_classification
Pichia
Pichia pastoris
Secretory protein
Biochemistry
Gene Expression Regulation, Fungal
Protein Biosynthesis
Chaperone (protein)
biology.protein
Secretion
Chemical chaperone
Glycoproteins
Biotechnology
Subjects
Details
- ISSN :
- 87567938
- Volume :
- 22
- Database :
- OpenAIRE
- Journal :
- Biotechnology Progress
- Accession number :
- edsair.doi.dedup.....b26b5aafb96a5b97af92f6ac67c96092