title: Engineering of vesicle trafficking improves heterologous protein secretion in Saccharomyces cerevisiae.
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.
authors.affiliation: Department of Chemical and Biological Engineering, Chalmers University of Technology, Göteborg, Sweden.
authors.forename: Jin
authors.initials: J
authors.lastname: Hou
authors.forename: Keith
authors.initials: K
authors.lastname: Tyo
authors.forename: Zihe
authors.initials: Z
authors.lastname: Liu
authors.forename: Dina
authors.initials: D
authors.lastname: Petranovic
authors.forename: Jens
authors.initials: J
authors.lastname: Nielsen
mesh_terms: D001692:Biological Transport / Q000235:genetics; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000378:metabolism; D006056:Golgi Apparatus / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000378:metabolism*; D000078790:Insulin Secretion; D060847:Metabolic Engineering / Q000379:methods; D051938:Munc18 Proteins / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D050600:SNARE Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000235:genetics; D022142:Secretory Vesicles / Q000235:genetics / Q000378:metabolism*; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism*
chemical_list: D007328:Insulin; D051938:Munc18 Proteins; D011994:Recombinant Proteins; C497931:SEC1 protein, S cerevisiae; C072520:SLY1 protein, S cerevisiae; D050600:SNARE Proteins; D029701:Saccharomyces cerevisiae Proteins; D000516:alpha-Amylases