ISHS Acta Horticulturae 459: International Symposium Brassica 97, Xth Crucifer Genetics Workshop
FROM FARMING FOR FOOD TO PHARMING FOR PHARMACEUTICALS AND OTHER HIGH VALUE PROTEINS IN CRUCIFERS |
| Authors: | G.J.H. van Rooijen, B. Kühnel, V. Kumar, J.-H. Liu, S. Mahmoud, M.M. Moloney |
| Keywords: | Oleosin, fusion, seed, enzyme, immobilization |
| Abstract:
Our laboratory has developed a novel strategy for the production and purification of foreign proteins and peptides in oilseeds. The strategy involves the fusion of the protein/peptide of interest to oleosin. Oleosins are structural proteins tightly associated with the oil body, the natural oil storage organelle of the plant seed. Proteins/peptides of interest could include sequences rich in essential amino acids, enzymes and therapeutic/pharmaceutical proteins. Oil bodies, and proteins associated with them, can be easily separated from the majority of other seed cell components using floatation separation/centrifugation, facilitating the purification of the desired protein. This technology was developed in canola (Brassica napus) for several reasons: it is amenable to genetic transformation, has a relatively high oil (body) content and is a widely grown crop plant. We are using this technology for several applications. One application is to improve the nutritional quality of the canola meal by expressing a nutritional peptide as an oleosin fusion. Using this technique we have successfully expressed a biologically active fish growth hormone on the surface of the oil body. A second application of this technology is to produce an oil body - oleosin-enzyme complex which functions as an immobilized catalyst. A third application is the production of an oleosin-fusion and the subsequent recovery of the recombinant fusion. Using this application we have successfully produced and purified a biologically active blood anti-coagulant hirudin. Oleosins accumulate in all seeds and it is likely that the production and recovery of oleosin fusion proteins could be done in all seeds. Therefore we are currently expanding this technology for use in several other crucifers such as Brassica carinata and Sinapis alba, and other oil seeds such as Linum usitatissimum L. (flax). | |
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