Study On Purification And Application Basis Of Chymopapain

Chymopapain is one of the most abundant enzymes in the crude papain, which has milk clotting and proteolytic activities. Chymopapain has very important application in hyperosteogeny treatment, identification of Rh blood group, accessory therapy of cancer, inflammatory disease treatment, cell separation. It is very difficult to isolate chymopapain from the latex of Carica papaya because chymopapain is very similar to the other three cysteine proteinases in the latex at the structure and properties . The isolation steps were complex and only can be used in small scale although pure chymopapain can be obtained currently. In this research, Chymopapain was purified by ion-exchange chromatography , ultrafitrate to concentrate, then vacuum freeze drying to save the sample. These simplified separation steps are very important to chymopapain preparation in large scale.Papain was used as adjuvant remedial agent in the course of radiotherapy and chemotherapy of cancer. The chymopapain has the higher pharmacological value than papain as it has more superiority in construction and physicochemical characterization than papain.The cytotoxicity of papain, chymopapain and five chemotherapeutic agents, which are Fluorouracil (5-Fu), Methotrexate(MTX), pingyangmycin(PYM), Vincristine(VCR) and Carboplatin(CBP). Chymopapain alone or together were observed by MTT method when the mouse ascites hepatoma cell line H22 cultured in different material concentration and different time in vitro in this study. The cytotoxicivity of chymopapain is higher than papain on the H22 cells with concentration dependency. Cooperation effect of chymopapain with five chemotherapeutic agents have concentration dependency. Chymopapain showed enhanced effect when added with five chemotherapeutic agents separately. The enhanced effect of chymopapain to drugs was the max when cultured on 48 hour.The cytotoxicities were measured by MTT method when human hepatoma cell line BEL-7402 cells were treated by different concentration chymopapain and 5-FU or MTX alone or together. The results are: The cytotoxicity of chymopapain on BEL-7402 showed concentration dependency. The cytotoxicity of chymopapain and combining 5-FU or MTX are more effective than 5-FU or MTX used alone on BEL-7402, when the non-cytotoxicity concentration of chymopapain was chosed. That is: chymopapain enhance cytotoxicity of 5-FU or MTX on BEL-7402 cells.