Research Paper | Published:

Enkephalins Prouduced in Transgenic Plants Using Modified 2S Seed Storage Proteins

Bio/Technologyvolume 7pages929932 (1989) | Download Citation



We have explored the possibility of producing large amounts of biologically active peptides as part of chimeric plant seed storage proteins, the 2S albumins. A portion of an Arabidopsis thaliana 2S albumin gene, encoding a region of the protein whose sequence is not highly conserved among different species, was replaced by sequences encoding the neuropeptide Leu-enkephalin, flanked by tryptic cleavage sites. Using Agrobacterium mediated transformation, the modified gene was transformed into Arabidopsis and oilseed rape and the 2S albumins isolated from the seeds of the regenerated transgenic plants. These were digested with trypsin and the enkephalin containing peptide isolated and treated to remove the extra lysine residue. Up to 200 nmol of peptide were recovered per gram of seed.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1

    Meienhofer, J. 1984. Large scale peptide synthesis: A review, p. 19–34. In: Peptides, Rag Marsson, U. (Ed.). Almquist & Wiksell International, Stockholm, Sweden.

  2. 2

    Josephson, S. and Bishop, R. 1988. Secretion of peptides from E coli: a production system for the pharmaceutical industry. Trends in Biotech. 6:218–224.

  3. 3

    Youle, R. and Huang, A.H.C. Occurrence of low molecular weight and high cysteine containing albumin storage protein in oilseeds of diverse species. Americ. J. Bot. 68:44–48.

  4. 4

    Lonnerdahl, B. and Janson, J.-C. 1972. Studies on Brassica seed protein. I. The low molecular weight proteins in rapeseed. Isolation and characterization. Biochim Biophys. Acta 278:175–183.

  5. 5

    Ampe, C., Van Damme, J., de Castro, L., Sampaio, M.J., Van Montagu, M., and Vandekerckhove, J. 1986. The amino-acid sequence of the 2S sulfur-rich proteins from seeds of Brazil nut (Bertholletia excelsa H.B.K.). Eur. J. Biochem. 159:597–604.

  6. 6

    Crouch, M.L., Tenbarge, K.M., Simon, A.E. and Ferl, R. 1983. cDNA clones for Brassica napus seed proteins: evidence from nucleotide sequence analysis that both subunits of napin are cleaved from a precursor polypeptide. J. Mol. Appl. Gen. 2:273–283.

  7. 7

    Ericson, M.L., Rodin, J., Lenman, M., Glimelius, K., Josefsson, L.-G. and Rak, L. 1986. Structure of the rapeseed 1.7S storage protein, napin, and its precursor. J. Biol. Chem. 261:14576–14581.

  8. 8

    Krebbers, E., Herdies, L., De Clercq, A., Seurinck, J., Leemans, J., Van Damme, J., Segura, M., Gheysen, G., Van Montagu, M. and Vandekerckhove, J. 1988. Determination of the processing sites of an Arabidopsis 2S albumin and characterization of the complete gene family. Plant Physiol. 87:859–866.

  9. 9

    Scofield, S.R. and Crouch, M.L. 1987. Nucleotide sequence of a member of the napin storage protein family from Brassica napus. J. Biol. Chem. 262:12202–12208.

  10. 10

    Josefsson, L.-G., Lenman, M., Ericson, M.L. and Rask, L. 1987. Structure of a gene encoding the 1.7S storage protein, napin, from Brassica napus. J. Biol. Chem. 262:12196–12201.

  11. 11

    Chrispeels, M.J., Higgins, J.V. and Spencer, D. 1982. Assembly of storage protein oligomers in the endoplasmic reticulum and processing of the polypeptides in the protein bodies of developing pea cotyledons. J. Cell Biology 93:306–313.

  12. 12

    Hughes, J., Smith, T.W., Klosterlitz, H.W., Fothergil, L.A., Morgan, B.A. and Morris, H.P. 1975. Identification of two related pentapeptides from the brain with potent opiate agonist activity. Nature 258:577–579.

  13. 13

    Hughes, J. 1975. Isolation of an endogenous compound from the brain with pharmacological properties similar to morphine. Brain Res 88:295–308.

  14. 14

    Duffaud, G. and Inouye, M. 1988. Signal peptidases recognize a structural feature at the cleavage site of secretory proteins. J. Biol. Chem. 263:10224–10228.

  15. 15

    Garnier, J., Asgutkorpe, D.J. and Robson, B. 1978. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J. Mol. Biol. 120:97–120.

  16. 16

    Sharief, F.S. and Li, S.S.-L. 1982. Amino acid sequence of small and large subunits of seed storage protein from Ricinus communis. J. Biol. Chem. 257:14753–14759.

  17. 17

    Seiringer, B.R., Liebisch, D.C., Gramsch, C., Herz, A., Weber, E., Evans, C.J., Esch, F.S. and Boehlen, P. 1985. Isolation and structure of a novel C-terminally amidated opioid peptide, amidorphin, from bovine adrenal medulla. Nature 313:57–59.

  18. 18

    Velten, J., Velten, L., Hain, R. and Schell, J. 1984. Isolation of a dual plant promoter fragment from the Ti plasmid of Agrobacterium tumefaciens. EMBO J. 3:2723–2730.

  19. 19

    Botterman, J. and Zabeau, M. 1987. A standardized vector system for manipulation and enhanced expression of genes in Escherichia coli. DNA 6:583–591.

  20. 20

    Cornelissen, M. and Vandewiele, M. 1989. Nuclear transcriptional activity of the tobacco plastid pbsA promoter. Nucleic Acids Res. 17:19–29.

  21. 21

    Deblaere, R., Reynaerts, A., Hofte, H., Hernalsteens, J.-P., Leemans, J. and Van Montagu, M. 1987. Vectors for cloning in plant cells. Methods in Enzymol. 153:277–29.

  22. 22

    Van Lijsebettens, M., Valvekens, D., Vanderhaeghen, H. and Van Montagu, M. 1989. Highly efficient Agrobacterium-mediated transformation of Arabidopsis thaliana: a genetic and molecular evaluation. In: Genetic manipulation in plant breeding, C. J. Jensen (Ed.). Plenum Press, New York (in press).

  23. 23

    Bidlingmeyer, B.A., Cohen, J.A. and Tarvin, T.L. 1984. Rapid analysis of amino acids using pre-column derivatization. J. Chromatogr. 336:93–104.

  24. 24

    Hirs, C.H.W. 1967. Detection of peptides by chemical methods. Methods in Enzymol. 11:325–329.

Download references

Author information


  1. Laboratorium voor Genetika, Rijksuniversiteit Gent, Ledeganckstraat 35, B-9000, Gent, Belgium

    • Joel Vandekerckhove
    • , Mieke Van Lijsebettens
    • , Martine Vandewiele
    • , Ann De Clercq
    •  & Marc Van Montagu
  2. Plant Genetic Systems N.V., J. Plateaustraat 22, B-9000, Gent, Belgium

    • Jozef Van Damme
    • , Johan Botterman
    • , Marc De Block
    • , Jan Leemans
    •  & Enno Krebbers


  1. Search for Joel Vandekerckhove in:

  2. Search for Jozef Van Damme in:

  3. Search for Mieke Van Lijsebettens in:

  4. Search for Johan Botterman in:

  5. Search for Marc De Block in:

  6. Search for Martine Vandewiele in:

  7. Search for Ann De Clercq in:

  8. Search for Jan Leemans in:

  9. Search for Marc Van Montagu in:

  10. Search for Enno Krebbers in:

About this article

Publication history



Issue Date


Further reading