title: Yeast mutant with efficient secretion identified by a novel secretory reporter, Cluc.
abstract: Yeast is an important host for the production of pharmaceutical or industrial proteins by virtue of its genetic information and easy handling. A number of heterologous proteins have been produced and purified from yeast cell cultures as secreted forms. Here, we describe a novel screening system of Saccharomyces cerevisiae and its application to improve the secretion efficiency of yeast. In our system, a natural secretory luciferase from Cypridina noctiluca is used as a reporter enzyme. The accumulation of enzymatically active luciferase in culture medium makes it possible to screen many samples simultaneously in a simple and sensitive assay. Using this system, we have discovered that the deletion mutant of MON2, which encoded a scaffold protein for vesicle formation located at the late Golgi, secreted luciferase highly efficiently to the extracellular space. Thus, we conclude that this new reporter assay is useful for the improvement and screening of yeast secretory strains.
authors.affiliation: Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology, 2-17-2-1 Tsukisamu-Higashi, Sapporo, Japan. nk-kanjou@aist.go.jp
authors.forename: Naoko
authors.initials: N
authors.lastname: Kanjou
authors.forename: Aki
authors.initials: A
authors.lastname: Nagao
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ohmiya
authors.forename: Satoru
authors.initials: S
authors.lastname: Ohgiya
mesh_terms: D000818:Animals; D003530:Cyprinidae / Q000378:metabolism*; D017930:Genes, Reporter / Q000235:genetics; D024861:Genetic Enhancement / Q000379:methods*; D008156:Luciferases / Q000096:biosynthesis* / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D008156:Luciferases