title: Production of Biologically Active Cecropin A Peptide in Rice Seed Oil Bodies.
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.
authors.affiliation: Institute of Food and Agricultural Technology-CIDSAV-XaRTA, University of Girona, Girona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Montesinos
authors.affiliation: Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Edifici CRAG, Campus de la UAB, Bellaterra, Barcelona, Spain.
authors.forename: Mireia
authors.initials: M
authors.lastname: Bundó
authors.affiliation: Mass Spectrometry Proteomics Platform, Biochimie et Physiologie Moléculaire des Plantes, SupAgro/INRA/CNRS/UMII/UMR 5004, Montpellier, France.
authors.forename: Esther
authors.initials: E
authors.lastname: Izquierdo
authors.affiliation: Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Edifici CRAG, Campus de la UAB, Bellaterra, Barcelona, Spain.
authors.forename: Sonia
authors.initials: S
authors.lastname: Campo
authors.affiliation: Institute of Food and Agricultural Technology-CIDSAV-XaRTA, University of Girona, Girona, Spain.
authors.forename: Esther
authors.initials: E
authors.lastname: Badosa
authors.affiliation: Mass Spectrometry Proteomics Platform, Biochimie et Physiologie Moléculaire des Plantes, SupAgro/INRA/CNRS/UMII/UMR 5004, Montpellier, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Rossignol
authors.affiliation: Institute of Food and Agricultural Technology-CIDSAV-XaRTA, University of Girona, Girona, Spain.
authors.forename: Emilio
authors.initials: E
authors.lastname: Montesinos
authors.affiliation: Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Edifici CRAG, Campus de la UAB, Bellaterra, Barcelona, Spain.
authors.forename: Blanca
authors.initials: B
authors.lastname: San Segundo
authors.affiliation: Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Edifici CRAG, Campus de la UAB, Bellaterra, Barcelona, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Coca
mesh_terms: D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000235:genetics; D018745:Genome, Plant; D066292:Lipid Droplets / Q000737:chemistry*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D010938:Plant Oils / Q000737:chemistry; D010940:Plant Proteins / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012639:Seeds / Q000235:genetics / Q000378:metabolism*; D019076:Transgenes
chemical_list: D023181:Antimicrobial Cationic Peptides; D010938:Plant Oils; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C102736:oleosin protein, Oryza sativa; C031161:cecropin A