Abstract
The yeast Saccharomyces cerevisiae is a widely used platform for the production of heterologous proteins of medical or industrial interest. However, heterologous protein productivity is often restricted due to the limitations of the host strain. In the protein secretory pathway, the protein trafficking between different organelles is catalyzed by the soluble NSF (N-ethylmaleimide-sensitive factor) receptor (SNARE) complex and regulated by the Sec1/Munc18 (SM) proteins. In this study, we report that over-expression of the SM protein encoding genes SEC1 and SLY1, improves the protein secretion in S. cerevisiae. Engineering Sec1p, the SM protein that is involved in vesicle trafficking from Golgi to cell membrane, improves the secretion of heterologous proteins human insulin precursor and α-amylase, and also the secretion of an endogenous protein invertase. Enhancing Sly1p, the SM protein regulating the vesicle fusion from endoplasmic reticulum (ER) to Golgi, increases α-amylase production only. Our study demonstrates that strengthening the protein trafficking in ER-to-Golgi and Golgi-to-plasma membrane process is a novel secretory engineering strategy for improving heterologous protein production in S. cerevisiae.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 120-127 |
| Number of pages | 8 |
| Journal | Metabolic Engineering |
| Volume | 14 |
| Issue number | 2 |
| DOIs | |
| State | Published - Mar 2012 |
Funding
We thank The Knut and Alice Wallenberg Foundation, EU Framework VII project SYSINBIO (Grant no. 212766 ), European Research Council project INSYSBIO (Grant no. 247013 ), the Chalmers Foundation, and NIH F32 Kirschstein NRSA fellowship for funding support.
Keywords
- Heterologous protein secretion
- Saccharomyces cerevisiae
- Sec1p
- Sly1p
- Vesicle trafficking
ASJC Scopus subject areas
- Applied Microbiology and Biotechnology
- Bioengineering
- Biotechnology