title: High-yield production of the VP1 structural protein epitope from serotype O foot-and-mouth disease virus in Escherichia coli.
abstract: For effective control of foot-and-mouth disease (FMD), the development of rapid diagnostic systems and vaccines are required against its etiological agent, FMD virus (FMDV). To accomplish this, efficient large-scale expression of the FMDV VP1 protein, with high solubility, needs to be optimized. We attempted to produce high levels of a serotype O FMDV VP1 epitope in Escherichia coli. We identified the subtype-independent serotype O FMDV VP1 epitope sequence and used it to construct a glutathione S-transferase (GST) fusion protein. For efficient production of the FMDV VP1 epitope fused to GST (VP1e-GST), four E. coli strains and three temperatures were examined. The conditions yielding the greatest level of VP1e-GST with highest solubility were achieved with E. coli BL21(DE3) at 25 °C. For high-level production, fed-batch cultures were conducted in 5-l bioreactors. When cells were induced at a high density and complex feeding solutions were supplied, approximately 11 g of VP1e-GST was obtained from a 2.9-l culture. Following purification, the VP1 epitope was used to immunize rabbits, and we confirmed that it induced an immune response.
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, Daejeon, Republic of Korea.
authors.forename: Joon-Goo
authors.initials: JG
authors.lastname: Jung
authors.forename: Yong Jae
authors.initials: YJ
authors.lastname: Lee
authors.forename: Natarajan
authors.initials: N
authors.lastname: Velmurugan
authors.forename: Young-Joon
authors.initials: YJ
authors.lastname: Ko
authors.forename: Hyang-Sim
authors.initials: HS
authors.lastname: Lee
authors.forename: Ki Jun
authors.initials: KJ
authors.lastname: Jeong
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D001483:Base Sequence; D061249:Batch Cell Culture Techniques; D019149:Bioreactors; D036022:Capsid Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D000939:Epitopes / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005536:Foot-and-Mouth Disease / Q000276:immunology / Q000517:prevention & control / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000737:chemistry / Q000145:classification / Q000235:genetics* / Q000276:immunology*; D005982:Glutathione Transferase / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D007114:Immunization; D008969:Molecular Sequence Data; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D015203:Reproducibility of Results; D012703:Serotyping; D014765:Viral Vaccines / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D000939:Epitopes; D011993:Recombinant Fusion Proteins; C442501:VP1 protein, Foot-and-mouth disease virus; D014765:Viral Vaccines; D005982:Glutathione Transferase