| Every kind of living creature has all superoxide dismutase. Now, superoxide dismutase has been isolated from germs, algae, fungi, high plant, insect, fish, birds etc. and applied to the daily life, such as the cosmetics, medicine, food, morbib examination of persons, animals and plants. But it is mostly abstracted from animals, because the superoxide dismutase content in animal is more than that in plant and the activity of the superoxide dismutase in animal is higher than that in plant. It is reported that aloe has many functions, such as eliminating inflammation, killing the viruses etc. The medicinal goods, cosmetics, healthful food have sold aloe. Till now, there is no report on the research that SOD is abstracted from aloe. According to above three aspects, the thesis studied the method in which SOD is abstracted from aloe and the natures of SOD, in order to understand completely the elements of aloe and establish base for the application and gene colony.The result is as follows: 1 Separate and purifyFirst, cut the clean leaves of the aloe into pieces and put Tris-HCl buffer cob in and mash up, abstract it and highly centrifuge. Secondly, put acetone in the abstracting liquor, put it aside for one hour and slowly centrifuge. Thirdly the deposit is fully dissolved and heated for 15minutes in 50 C water, and get crude enzyme solution through centrifuge. Fourthly, according to the activity, collect, dialyse , freeze, dry respectively the SOD protein through DEAE-sepharose column chromatog - raphy ; Process the SOD protein through Sephacryl S-200 column chromatography with the preceding method. At last, process the pure SOD protein with same functional enzyme electrophoresis and active dye, isoelectric focusing electrophoresis and SDS-PAGE. The result is as following: SOD-I, SOD-II, SOD-IIIare completely purified. Recovery of SOD-I is 3. 4% and purified times of SOD- I is 60. 0; that of SOD-II is 5. 6%and that of SOD-II is 153; that of SOD-IIIis 4.2% and that of SOD-III is 84.6. In this case, recovery and purified times of SOD-IIare the highest in the three same functional enzymes.2 Nature studyCu. Zn- SOD is sensitive to KCN and H2O2. Mn-SOD is not sensitive to KCN and H2O2. Fe-SOD is sensitive to H2O2. but not sensitive to KCN. 1mmo1/LKCN is not influential to the activity of SOD. 5mmol/L H2O2 can completely kill the three kinds of enzymes .So we can come to the conclusion that SOD in aloe is Fe-SOD.Determined by Sephacryl S-200 column, the molecular weight of SOD-I is approximately 46 000D, that of SOD-II is 35 000D, that of SOD-IIIis 40000D. Determined by SDD-PAGE, the subunit molecular weight of SOD- I isapproximately 23 000D; that of SOD-II is 17 000D; that of SOD-IIIisl9 000D.Therefore, SOD in aloe is all Fe-SOD and are made of two same subunits.Isoelectric point of the three kinds of pur SOD are following: pI of SOD-I is 8.9; pI of SOD-IIis 7.6; pI of SOD-ffl is 7.35.Some chemical modifications suchas PMSF, DTT, PCMB, NBS, BrAc, IAc, SUAN, TNBS is not fit to SOD...
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