title: Versatile signal peptide of Flavobacterium-originated organophosphorus hydrolase for efficient periplasmic translocation of heterologous proteins in Escherichia coli.
abstract: Organophosphorus hydrolase (OPH) from Flavobacterium species is a membrane-associated homodimeric metalloenzyme and has its own signal peptide in its N-terminus. We found that OPH was translocated into the periplasmic space when the original signal peptide-containing OPH was expressed in recombinant Escherichia coli even though its translocation efficiency was relatively low. To investigate the usability of this OPH signal peptide for periplasmic expression of heterologous proteins in an E. coli system, we employed green fluorescent protein (GFP) as a cytoplasmic folding reporter and alkaline phosphatase (ALP) as a periplasmic folding reporter. We found that the OPH signal peptide was able to use both twin-arginine translocation (Tat) and general secretory (Sec) machineries by switching translocation pathways according to the nature of target proteins in E. coli. These results might be due to the lack of Sec-avoidance sequence in the c-region and a moderate hydrophobicity of the OPH signal peptide. Interestingly, the OPH signal peptide considerably enhanced the translocation efficiencies for both GFP and ALP compared with commonly used TorA and PelB signal peptides that have Tat and Sec pathway dependences, respectively. Therefore, this OPH signal peptide could be successfully used in recombinant E. coli system for efficient periplasmic production of target protein regardless of the subcellular localization where functional folding of the protein occurs. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:848-854, 2016.
authors.affiliation: Dept. of Chemical Engineering, Pohang University of Science and Technology, Pohang, 790-784, Korea.
authors.forename: Dong Gyun
authors.initials: DG
authors.lastname: Kang
authors.affiliation: School of Chemical Engineering, Yeungnam University, Gyeongsan, 712-749, Korea.
authors.forename: Jeong Hyun
authors.initials: JH
authors.lastname: Seo
authors.affiliation: Dept. of Chemical Engineering, Pohang University of Science and Technology, Pohang, 790-784, Korea.
authors.forename: Byung Hoon
authors.initials: BH
authors.lastname: Jo
authors.affiliation: School of Biotechnolgy and Graduate School of Biochemistry, Yeungnam University, Gyeongsan, 712-749, Korea.
authors.forename: Chang Sup
authors.initials: CS
authors.lastname: Kim
authors.affiliation: Dept. of Biological and Environmental Engineering, Semyung University, Jecheon, 390-711, Korea.
authors.forename: Suk Soon
authors.initials: SS
authors.lastname: Choi
authors.affiliation: Dept. of Chemical Engineering, Pohang University of Science and Technology, Pohang, 790-784, Korea.
authors.forename: Hyung Joon
authors.initials: HJ
authors.lastname: Cha
mesh_terms: D043303:Aryldialkylphosphatase / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology / Q000378:metabolism*; D005417:Flavobacterium / Q000201:enzymology*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D019897:Periplasm / Q000737:chemistry / Q000378:metabolism*
chemical_list: D010455:Peptides; D043303:Aryldialkylphosphatase
keywords: Escherichia coli; heterologous protein; organophosphorous hydrolase; periplasmic translocation; signal peptide