title: A novel fusion partner for enhanced secretion of recombinant proteins in Saccharomyces cerevisiae.
abstract: Expressing proteins with fusion partners improves yield and simplifies the purification process. We developed a novel fusion partner to improve the secretion of heterologous proteins that are otherwise poorly excreted in yeast. The VOA1 (YGR106C) gene of Saccharomyces cerevisiae encodes a subunit of vacuolar ATPase. We found that C-terminally truncated Voa1p was highly secreted into the culture medium, even when fused with rarely secreted heterologous proteins such as human interleukin-2 (hIL-2). Deletion mapping of C-terminally truncated Voa1p, identified a hydrophilic 28-amino acid peptide (HL peptide) that was responsible for the enhanced secretion of target protein. A purification tag and a protease cleavage site were added to use HL peptide as a multi-purpose fusion partner. The utility of this system was tested via the expression and purification of various heterologous proteins. In many cases, the yield of target proteins fused with the peptide was significantly increased, and fusion proteins could be directly purified with affinity chromatography. The fusion partner was removed by in vitro processing, and intact proteins were purified by re-application of samples to affinity chromatography.
authors.affiliation: Cell Factory Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 34141, Republic of Korea.
authors.forename: Jung-Hoon
authors.initials: JH
authors.lastname: Bae
authors.affiliation: Cell Factory Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 34141, Republic of Korea.
authors.forename: Bong Hyun
authors.initials: BH
authors.lastname: Sung
authors.affiliation: Industrial Microbiology and Bioprocess Research Center, Korean Research Institute of Bioscience and Biotechnology, Jeongeup, 56212, Republic of Korea.
authors.forename: Jeong-Woo
authors.initials: JW
authors.lastname: Seo
authors.affiliation: Industrial Microbiology and Bioprocess Research Center, Korean Research Institute of Bioscience and Biotechnology, Jeongeup, 56212, Republic of Korea.
authors.forename: Chul Ho
authors.initials: CH
authors.lastname: Kim
authors.affiliation: Cell Factory Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 34141, Republic of Korea. sohn4090@kribb.re.kr.
authors.forename: Jung-Hoon
authors.initials: JH
authors.lastname: Sohn
mesh_terms: D007376:Interleukin-2 / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism*
chemical_list: C508594:IL2 protein, human; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C000613612:VOA1 protein, S cerevisiae; D025262:Vacuolar Proton-Translocating ATPases
keywords: Fusion partner; Purification; Saccharomyces cerevisiae; VOA1