Production of Biologically Active Cecropin A Peptide in Rice Seed Oil Bodies

PLoS One. 2016 Jan 13;11(1):e0146919. doi: 10.1371/journal.pone.0146919. eCollection 2016.

Abstract

Cecropin A is a natural antimicrobial peptide that exhibits fast and potent activity against a broad spectrum of pathogens and neoplastic cells, and that has important biotechnological applications. However, cecropin A exploitation, as for other antimicrobial peptides, is limited by their production and purification costs. Here, we report the efficient production of this bioactive peptide in rice bran using the rice oleosin 18 as a carrier protein. High cecropin A levels were reached in rice seeds driving the expression of the chimeric gene by the strong embryo-specific oleosin 18 own promoter, and targeting the peptide to the oil body organelle as an oleosin 18-cecropin A fusion protein. The accumulation of cecropin A in oil bodies had no deleterious effects on seed viability and seedling growth, as well as on seed yield. We also show that biologically active cecropin A can be easily purified from the transgenic rice seeds by homogenization and simple flotation centrifugation methods. Our results demonstrate that the oleosin fusion technology is suitable for the production of cecropin A in rice seeds, which can potentially be extended to other antimicrobial peptides to assist their exploitation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antimicrobial Cationic Peptides / biosynthesis*
  • Antimicrobial Cationic Peptides / genetics
  • Genome, Plant
  • Lipid Droplets / chemistry*
  • Mass Spectrometry
  • Molecular Sequence Data
  • Oryza / genetics
  • Oryza / metabolism*
  • Phenotype
  • Plant Oils / chemistry
  • Plant Proteins / metabolism
  • Plants, Genetically Modified / metabolism
  • Promoter Regions, Genetic
  • Recombinant Fusion Proteins / metabolism
  • Seeds / genetics
  • Seeds / metabolism*
  • Transgenes

Substances

  • Antimicrobial Cationic Peptides
  • Plant Oils
  • Plant Proteins
  • Recombinant Fusion Proteins
  • oleosin protein, Oryza sativa
  • cecropin A

Grants and funding

This work was supported by the Spanish Ministry of Economy and Competitiveness [Grants EUI2008-03769, EUI2008-03572 and BIO2012-32838], and by the French Research Agency [Grant ANR-08-KBBE-010]. The funders had no role in study design, data collection and analysis, decisión to publish, or preparation of the manuscript.