꽃게 (P. trituberculatus) hemolymph를 채취한 후 원심분리 및 DEAE-Sephadex A-50 column chromatgraphy를 행하여 hemocyanin을 정제하였다. 정제된 hemocyanin은 단일 polypeptide로 이루어져 있었으며 monomer의 분자량은 약 70,000으로 추정 되었다. 또한 꽃게 hemolymph 단백질중 hemocyanin의 함량은 46%, Cu는 단일 polypeptide 당 2 mole, 그리고 당 함량은 단백질 mg 당 $10;{mu}g$ 이었다. 한편 정제된 hemocyanin은 Ca등 2가 이온을 가하지 않고 pH 5.7~8.9인 상태에서 mono, di, tri, tetra, pentamer등 다양한 polymorphism을 나타내었다.
The hemocyanin from the hemolymph of crab, Portunus trituberculatus which belong to crustacean was purified by a single step of ion-exchange column chromatography after dialysis and ultracentrifugation. The purified hemocyanin was revealed a high degree of polymorphism after analyzed it by Davis-polyacrylamide gel electrophoresis, the peroxidase activity, and copper staining method. On the other hand, the denatured hemocyanin was appeared to be homogeneous polypeptide by SDS-PAGE, and the approximate molecular weight was estimated to be 70,000. It was found that the purified hemocyanin was a glycoprotein in nature that contained 2 moles of copper/l mole of hemocyanin, and it was estimated that their contents in normal body fluid were approximately 46%.
