Plant seed oil-bodies as carriers for foreign proteins.
Plant seeds frequently store oils (triglycerides) in discrete organelles called oil-bodies. These are normally surrounded by a phospholipid half-unit membrane equipped with specialized proteins called oleosins. Oleosins are highly lipophilic proteins, are expressed at high levels in many seeds and are specifically targeted to oil-bodies. We have investigated the potential of oleosins to act as carriers for recombinant proteins by the production of translational fusions between oleosins and genes encoding proteins foreign to plant cells. We have shown that a fusion comprising a complete oleosin coding domain and a beta-glucuronidase coding sequence may be expressed specifically in the seeds of the oilseed crop plant, Brassica napus, and its product is correctly targeted with approximately 80% of the activity partitioning with oil-bodies. Recombinant oil-bodies may be used to facilitate separation of a recombinant protein from other cellular proteins. Using this approach, the desired protein may be cleaved from the oil-bodies using an endoprotease and further purified. Alternatively, a fusion protein which is enzymatically active and resides on the oil-bodies may be used directly in heterogeneous catalysis. In this application, after a round of catalysis the oil-bodies may be recovered and re-used several times without loss of activity. Thus the oil-bodies act as an immobilization matrix. The fusion protein is stable in dry seeds for long periods and when extracted has a half-life of 3-4 weeks on oil-bodies. Finally, the production of these recombinant oil-bodies is extremely inexpensive, offering a novel route to the manufacture of recombinant proteins.
van Rooijen
G J
GJ
Department of Biological Sciences, University of Calgary, Alberta, Canada.
Moloney
M M
MM
D029681:Arabidopsis Proteins; D001937:Brassica; D002352:Carrier Proteins; D004355:Drug Stability; D005966:Glucuronidase / Q000235:genetics; D015388:Organelles / Q000378:metabolism*; D010938:Plant Oils / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins; D012639:Seeds / Q000648:ultrastructure*
D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D010938:Plant Oils; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C079353:oleosin protein, Arabidopsis; C079356:oleosin protein, Brassica napus; D005966:Glucuronidase