title: An Efficient Genome-Wide Fusion Partner Screening System for Secretion of Recombinant Proteins in Yeast.
abstract: To produce rarely secreted recombinant proteins in the yeast Saccharomyces cerevisiae, we developed a novel genome-wide optimal translational fusion partner (TFP) screening system that involves recruitment of an optimal secretion signal and fusion partner. A TFP library was constructed from a genomic and truncated cDNA library by using the invertase-based signal sequence trap technique. The efficiency of the system was demonstrated using two rarely secreted proteins, human interleukin (hIL)-2 and hIL-32. Optimal TFPs for secretion of hIL-2 and hIL-32 were easily selected, yielding secretion of these proteins up to hundreds of mg/L. Moreover, numerous uncovered yeast secretion signals and fusion partners were identified, leading to efficient secretion of various recombinant proteins. Selected TFPs were found to be useful for the hypersecretion of other recombinant proteins at yields of up to several g/L. This screening technique could provide new methods for the production of various types of difficult-to-express proteins.
authors.affiliation: Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea.
authors.forename: Jung-Hoon
authors.initials: JH
authors.lastname: Bae
authors.affiliation: Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea.
authors.forename: Bong Hyun
authors.initials: BH
authors.lastname: Sung
authors.affiliation: Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea.
authors.forename: Hyun-Jin
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea.
authors.forename: Soon-Ho
authors.initials: SH
authors.lastname: Park
authors.affiliation: Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea.
authors.forename: Kwang-Mook
authors.initials: KM
authors.lastname: Lim
authors.affiliation: Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea.
authors.forename: Mi-Jin
authors.initials: MJ
authors.lastname: Kim
authors.affiliation: 1] Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea [2] Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea.
authors.forename: Cho-Ryong
authors.initials: CR
authors.lastname: Lee
authors.affiliation: 1] Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea [2] Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea.
authors.forename: Jung-Hoon
authors.initials: JH
authors.lastname: Sohn
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D005285:Fermentation; D015723:Gene Library; D016681:Genome, Fungal; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism; D007378:Interleukins / Q000378:metabolism; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D043324:beta-Fructofuranosidase / Q000378:metabolism
chemical_list: C496971:IL32 protein, human; D007376:Interleukin-2; D007378:Interleukins; D011993:Recombinant Fusion Proteins; D043324:beta-Fructofuranosidase