Utilization of waste protein from oil seed by improvement for its quality

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The first objective of this study was determination of the crystal structure of cruciferin from rapeseed by x-ray crystallography. Since native cruciferin forms heterogenic hexamer composed of at least three kinds of subunit, it is actually difficult to make its crystal for x-ray crystallography. Thus, we constructed the E. coli expression system to prepare recombinant procruciferin. The cDNA was obtained after RT-PCR reaction using mRNA purified from its developing seeds. As a result of investigation for various expression conditions, the recombinant procruciferin occupied 15% of total E. coli protein as a soluble protein. Using this expression system, wild type procruciferin and some mutants were prepared to make crystals and characterize their physicochemical properties. The amino acid residues forming intra salt bridge and C287 residue were mutated. In addition, variable regions of the procruciferin were modified. Another objective was the production of engineered rape seed plant that expresses soybean glycinin in its seed. Because the glycinin has better functional and nutritional properties, we can expect the improvement by adding the better properties to the rape seed. The cotyledon of the rape seed was transformed by agrobacterium method, and the expression of glycinin in the seed was observed by western blotting. Eventually, flowering F1 generation was obtained.