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Moderate Expression of SEC16 Increases Protein Secretion by Saccharomyces cerevisiae

Jichen Bao (Institutionen för biologi och bioteknik, Systembiologi) ; Mingtao Huang (Institutionen för biologi och bioteknik, Systembiologi) ; Dina Petranovic (Institutionen för biologi och bioteknik, Systembiologi) ; Jens B. Nielsen (Institutionen för biologi och bioteknik, Systembiologi)
Applied and Environmental Microbiology (0099-2240). Vol. 83 (2017), 14, p. Article no. UNSP e03400-16 .
[Artikel, refereegranskad vetenskaplig]

The yeast Saccharomyces cerevisiae is widely used to produce biopharmaceutical proteins. However, the limited capacity of the secretory pathway may reduce its productivity. Here, we increased the secretion of a heterologous beta-amylase, a model protein used for studying the protein secretory pathway in yeast, by moderately overexpressing SEC16, which is involved in protein translocation from the endoplasmic reticulum to the Golgi apparatus. The moderate overexpression of SEC16 increased beta-amylase secretion by generating more endoplasmic reticulum exit sites. The production of reactive oxygen species resulting from the heterologous beta-amylase production was reduced. A genome-wide expression analysis indicated decreased endoplasmic reticulum stress in the strain that moderately overexpressed SEC16, which was consistent with a decreased volume of the endoplasmic reticulum. Additionally, fewer mitochondria were observed. Finally, the moderate overexpression of SEC16 was shown to improve the secretion of two other recombinant proteins, Trichoderma reesei endoglucanase I and Rhizopus oryzae glucan-1,4-beta-glucosidase, indicating that this mechanism is of general relevance. IMPORTANCE There is an increasing demand for recombinant proteins to be used as enzymes and pharmaceuticals. The yeast Saccharomyces cerevisiae is a cell factory that is widely used to produce recombinant proteins. Our study revealed that moderate overexpression of SEC16 increased recombinant protein secretion in S. cerevisiae. This new strategy can be combined with other targets to engineer cell factories to efficiently produce protein in the future.

Nyckelord: protein secretion, mitochondria, ERES, ROS, SEC16



Denna post skapades 2017-08-14. Senast ändrad 2017-09-06.
CPL Pubid: 251114

 

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Institutioner (Chalmers)

Institutionen för biologi och bioteknik, Systembiologi

Ämnesområden

Mikrobiologi

Chalmers infrastruktur

 


Projekt

Denna publikation är ett resultat av följande projekt:


Industrial Systems Biology of Yeast and A. oryzae (INSYSBIO) (EC/FP7/247013)