豬血中提取CuZn-SOD

生物工程論文編號:SW035  論文字數(shù):12803,頁數(shù):27

目  錄
中文摘要 I
英文摘要 II
目錄 III
1 緒論 1
2 實驗部分 3
2.1 實驗材料與試劑 3
2.2 實驗儀器設(shè)備 3
2.3 實驗步驟 4
2.4 Cu/Zn-SOD酶活的測定 6
2.5 Cu/Zn-SOD蛋白濃度的測定 7
3 結(jié)果與討論 9
3.1 實驗室制備條件的研究 9
3.2 工藝放大條件優(yōu)化及其制備 13
4 總結(jié)與展望 20
4.1 總結(jié) 20
4.2 展望 21
致謝 22
參考文獻 23

摘   要：超氧化物歧化酶(superoxide dismutase，SOD)是生物體內(nèi)重要的超氧陰離子自由基清除劑，能催化生物體內(nèi)超氧自由基(O2-)發(fā)生歧化反應(yīng)，對機體細胞起保護作用，己成為化學及生物化學研究領(lǐng)域中熱門的研究課題。本文對豬血中提取Cu/Zn-SOD的工藝條件進行了研究，首先在實驗室條件下從豬血中提取Cu/Zn-SOD，并對其離心條件，乙醇-氯仿處理后的混合液的處理，丙酮沉淀等提取條件進行了優(yōu)化。對實驗室條件下所得的工藝條件進行了初步放大實驗，工藝放大流程及實驗結(jié)果為：1000ml豬血經(jīng)過血液預(yù)處理，溶血，乙醇-氯仿混合液除去血紅蛋白，丙酮分級沉淀，60-65℃熱變性及丙酮二次沉淀得到粗Cu/Zn-SOD酶液。再將粗酶液上陰離子DEAE-Sepharose Fast Flow交換層析柱，凍干，最終獲得了比活力為5660.79U/mg的精品Cu/Zn-SOD酶23.96mg。
 
關(guān)鍵詞：豬血；銅鋅超氧化物歧化酶；提取；優(yōu)化

The Extraction And Properties Of Cu/Zn-SOD From Porcine  Blood

Abstract: Superoxide dismutase (SOD) plays a very important role in organisms from oxygen toxicity, which can catalyze oxygen toxicity to happen disproportionatio, it protects the organism cell, and has become of chemical and biochemical research in the field of hot research topics. This paper studies extraction from the blood of Cu/Zn-SOD process conditions, First in laboratory extracted from the blood of Cu/Zn-SOD, and the extraction conditions optimization, hence amplification process of extracting the best conditions. After optimizing the above-mentioned process flow, abtained Large engineering and technology results : porcine erytrocyte coarse Cu/Zn-SOD was obtained by several steps, which were about pretreatment of fresh blood hemolysis, removing hemoglobin by ethanol and chloroform, precipition by acetone, heat denaturation, and precipition by acetone again etc. Then, through DEAE-Sepharose Fast Flow, it was completely purified. The specific activity of SOD was 5660.79U/mg, and its weight was 23.96mg.
 
Key words: Porcine blood; Cu/Zn-SOD; Extract; Optimize