During the protein secretory pathway, an important step is the fusion of vesicles with target membranes, which is catalyzed by SNARE proteins. Sec1/Munc18 (SM) proteins are key regulators of vesicle docking and SNARE complex assembly. In this study, we show that engineering SM proteins Sly1p and Sec1p which regulate the vesicles fusion of ER-to-Golgi and Golgi-to-plasma membrane, respectively, enhanced the heterologous protein production in Saccharomyces cerevisiae. SLY1 overexpression increased human insulin precursor secretion by 10% and α-amylase by 43%. SEC1 overexpression improved human insulin precursor secretion by 28% and α-amylase by 62%. The results suggested that the protein secretion from Golgi to plasma membrane is a rate-limiting step for both insulin precursor and α-amylase production. While improving the vesicle trafficking from ER to Golgi enhances secretory capability of α-amylase, but cannot improve the insulin precursor very effectively. Our data shows that vesicle trafficking engineering provides an attractive strategy to improve heterologous protein secretion in Saccharomyces cerevisiae.
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Overexpression Vesicle-Trafficking Protein and the Impact to Heterologous Protein Production In Saccharomyces Cerevisiae
During the protein secretory pathway, an important step is the fusion of vesicles with target membranes, which is catalyzed by SNARE proteins. Sec1/Munc18 (SM) proteins are key regulators of vesicle docking and SNARE complex assembly. In this study, we show that engineering SM proteins Sly1p and Sec1p which regulate the vesicles fusion of ER-to-Golgi and Golgi-to-plasma membrane, respectively, enhanced the heterologous protein production in Saccharomyces cerevisiae. SLY1 overexpression increased human insulin precursor secretion by 10% and α-amylase by 43%. SEC1 overexpression improved human insulin precursor secretion by 28% and α-amylase by 62%. The results suggested that the protein secretion from Golgi to plasma membrane is a rate-limiting step for both insulin precursor and α-amylase production. While improving the vesicle trafficking from ER to Golgi enhances secretory capability of α-amylase, but cannot improve the insulin precursor very effectively. Our data shows that vesicle trafficking engineering provides an attractive strategy to improve heterologous protein secretion in Saccharomyces cerevisiae.