Extraction Of Superoxide Dismutase(SOD) And Heme From Porcine And The Application Of SOD

Pig blood is very rich in our country, it is rich in protein and amino acid is balance, it is the source of high-quality protein. Red blood cell is the principal components in pig blood, the main protein in red blood cell is hemoglobin, account for more than 90%. To develop pig blood resources has good economic and social benefits, it is not only can increase the protein of feed sources, increase raw material resources for food, medicine and other fields, but also to reduce the pollution of environment from animal waste. In this thesis, SOD and heme were extracted from pig blood, structural identification, stability in stored procedure of SOD and heme, the in vitro antioxidant activity and the effect on mouse-borne tumor cell apoptosis of SOD were studied.1 The optimization extraction condition of SOD and hemeWhen broken the cell membrane by ultrasonic, the optimum ultrasonic conditions for high SOD specific activity and high heme extraction rate were:ultrasonic power 70 W, ultrasonic time 15min and ultrasonic temperature 40℃; Using Box-Behnken of the central composite rotary design test, Design-Expert software to process data, the best extraction conditions obtained SOD:heating temperature 65.2℃, heating time 26.1 min, copper chloride solution of 0.1 mol/L added 0.423 mL/15 mL dissolve blood; the optimum extraction conditions of heme:acidic acetone addition level was 5.66 times that of hemoglobin, ratio of hydrochloric acid and acetone was 3%, the extraction time was 42.00 min, the maximum extraction rate of heme was 0.225 g/100 mL.2 Initial structure identification of SOD and hemeAnalyzed extracted SOD samples by UV spectrum, the maximum absorption peak is 265 nm, the result was consistent with those reported; compared the spectrogram of standard heme and samples heme by UV spectrum, Infrared spectrum and high performance liquid chromatography, the extraction sample was proved to be heme. Under the choosen mobile phase(methanol:water was 80:20) and equipment condition, detection limits, precision and stability was good, purity of heme was 72%. 3 Storage stability of SOD and hemeFreezing storage was conducive to the stability of SOD and heme in storage. In freezing storage, chill storage, room temperature and dark storage, room temperature in daylight storage, the relatively activity of SOD were 90%,85%,82%and 80.25%after 20 h. In freezing storage, chill storage, room temperature and dark storage, room temperature in daylight storage, the relatively contents of heme were 91.58%,88.61%,83.46% and 81.75% after 9 weeks.4 Antioxidant activity and effects on mouse-borne tumor sp2/0 cell apoptosis of SODThe antioxidant activity of SOD was detected in vitro, using Vc as the reference standard, the results showed that SOD had good antioxidant activity, but lower than that of Vc.Observation of cell morphology is the basic method of apoptosis study. It was indicated that mouse-borne tumor SP2/0 cell grew well without SOD, showing round; mouse-borne tumor SP2/0 cell was significant nuclear condensation, fragmentation, showing the apoptotic morphology with different SOD concentration after a while, and with the increasing of the concentration, the extent of nuclear condensation and fragmentation increased. SOD can inhibit mouse-borne tumor SP2/0 cell proliferation to some extent in vitro, and showed time-effect and dose-effect relationship, SOD could induce mouse-borne tumor SP2/0 cell apoptosis.