BioMetals

, Volume 26, Issue 1, pp 113–122 | Cite as

Expression and characterization of recombinant bovine lactoferrin in E. coli

  • Isui García-Montoya
  • Jose Salazar-Martínez
  • Sigifredo Arévalo-Gallegos
  • Sugey Sinagawa-García
  • Quintin Rascón-Cruz
Article
First Online:
Received:
Accepted:

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.

Keywords

Bovine lactoferrin Antibacterial activity Thrombin Fusion expression Affinity chromatography 
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Notes

Acknowledgments

This study was supported by internal resources found in the Universidad Autonoma de Chihuahua and Proteo/MuuTechnologies de Mexico.

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Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Isui García-Montoya
    • 1
  • Jose Salazar-Martínez
    • 2
  • Sigifredo Arévalo-Gallegos
    • 1
  • Sugey Sinagawa-García
    • 3
  • Quintin Rascón-Cruz
    • 1
  1. 1.Laboratorio de Biotecnología, Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua, Circuito 1Nuevo Campus UniversitarioChihuahuaMexico
  2. 2.Proteo/Muu-Technologies de MexicoGomez PalacioMexico
  3. 3.Facultad de Agronomía, Campus de Ciencias Agropecuarias, Universidad Autónoma de Nuevo LeónMonterreyMexico

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