title: Expression and characterization of recombinant bovine lactoferrin in E. coli.
abstract: Lactoferrin is a member of the transferrin family of iron-binding proteins with a number of properties, including antibacterial activity against a broad spectrum of Gram-negative and Gram-positive bacteria. bovine lactoferrin cDNA was isolated, cloned and expressed as a fusion protein. The amino acid sequence of the fusion was analyzed and compared with other species. Crystallographic data were used to compare structural differences between bovine and human lactoferrin in 3-D models. A thioredoxin fusion protein was expressed and shown to have a different molecular weight compared with native bLf. After purification using Ni-NTA, the yield of recombinant bovine lactoferrin was 15.3 mg/l with a purity of 90.3 %. Recombinant bLf and pepsin-digested rbLf peptides demonstrated antibacterial activity of 79.8 and 86.9 %, respectively. The successful expression of functional, active and intact rbLf allows us to study the biochemical interactions of antimicrobial proteins and peptides and will facilitate their study as immunomodulators.
authors.affiliation: Laboratorio de Biotecnología, Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua, Circuito 1, Nuevo Campus Universitario, 31125, Chihuahua, Mexico.
authors.forename: Isui
authors.initials: I
authors.lastname: García-Montoya
authors.forename: Susana Aideé
authors.initials: SA
authors.lastname: González-Chávez
authors.forename: Jose
authors.initials: J
authors.lastname: Salazar-Martínez
authors.forename: Sigifredo
authors.initials: S
authors.lastname: Arévalo-Gallegos
authors.forename: Sugey
authors.initials: S
authors.lastname: Sinagawa-García
authors.forename: Quintin
authors.initials: Q
authors.lastname: Rascón-Cruz
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D002417:Cattle; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000187:drug effects*; D015870:Gene Expression; D007781:Lactoferrin / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D040681:Structural Homology, Protein
chemical_list: D000900:Anti-Bacterial Agents; D011994:Recombinant Proteins; D007781:Lactoferrin