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Production of biologically active Cecropin A peptide in rice seed oil bodies

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

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]

Public Library of Science (PLoS)

Manager: Ministerio de Educación y Ciencia (Espanya)
Author: Montesinos Barreda, Laura
Bundó, Mireia
Izquierdo, Esther
Campo, Sonia
Badosa Romañó, Esther
Rossignol, Michel
Montesinos Seguí, Emilio
San Segundo de los Mozos, Blanca
Coca, María
Date: 2016 January 13
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
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]
Format: application/pdf
Document access: http://hdl.handle.net/10256/11910
Language: eng
Publisher: Public Library of Science (PLoS)
Collection: info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0146919
info:eu-repo/semantics/altIdentifier/eissn/1932-6203
info:eu-repo/semantics/altIdentifier/eissn/1932-6203
info:eu-repo/grantAgreement/MICINN//EUI2008-03572/ES/PLANTAS BIOFACTORIA DE PEPTIDOS ANTIMICROBIANOS/
Rights: Reconeixement 3.0 Espanya
Rights URI: http://creativecommons.org/licenses/by/3.0/es
Subject: Pèptids
Plantes transgèniques
Arròs
Peptides
Transgenic plants
Rice
Arròs -- Protecció
Rice -- Protection
Llavors -- Protecció
Seeds -- Protection
Title: Production of biologically active Cecropin A peptide in rice seed oil bodies
Type: info:eu-repo/semantics/article
Repository: DUGiDocs

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