通过电倾析法进行酶的纯化。
Enzyme purification by electrodecantation.
作者信息
Winchester B G, Caffrey M, Robinson D
出版信息
Biochem J. 1971 Jan;121(2):161-8. doi: 10.1042/bj1210161.
Abstract
- Electrodecantation has been applied to the initial fractionation from crude material of pig kidney beta-d-glucosidase and sheep testicular N-acetylglucosaminidase. 2. The isoelectric points determined by isoelectric focusing were pH4.9 for the beta-d-glucosidase and pH6.3 for the N-acetylglucosaminidase. 3. Electrodecantation of pig kidney extract and sheep testicular extract was carried out at pH4.9 and 6.3 respectively. 4. A six- to ten-fold increase in specific activity could be obtained with good recoveries after a single cycle of electrodecantation. 5. The technique has also been used to purify further an extracellular Bacillus subtilis protease preparation. 6. Attempts to use electrodecantation for the concentration of very dilute enzyme solutions resulted in considerable loss of activity. 7. The limitations and potential use of the technique in laboratory-scale enzyme preparation are discussed.
摘要
- 电倾析法已应用于从猪肾β - d - 葡萄糖苷酶和羊睾丸N - 乙酰葡糖胺酶的粗原料中进行初步分级分离。2. 通过等电聚焦测定的等电点，β - d - 葡萄糖苷酶为pH4.9，N - 乙酰葡糖胺酶为pH6.3。3. 猪肾提取物和羊睾丸提取物的电倾析分别在pH4.9和6.3下进行。4. 经过单个电倾析循环后，比活性可提高6至10倍，且回收率良好。5. 该技术还用于进一步纯化枯草芽孢杆菌细胞外蛋白酶制剂。6. 尝试用电倾析法浓缩非常稀的酶溶液导致相当大的活性损失。7. 讨论了该技术在实验室规模酶制剂制备中的局限性和潜在用途。
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Nature. 1962 Jul 21;195:281-3. doi: 10.1038/195281a0.
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