Sicard P J, Mialonier G, Smagghe M
Prep Biochem. 1978;8(1):19-36. doi: 10.1080/00327487808068216.
The complete purification of renin raises difficult problems due to its extremely low concentration in kidney (less than 1/50,000 of total proteins). The complete purification of hog kidney renin has been realized on a large scale, starting from 300 kg of fresh hog kidneys. 14.6 mg of pure renin were obtained with an overall yield of 4%. The purification procedure involved 14 steps. The enzyme was extracted at pH 3.5. Subsequent purification steps were performed in the presence of protease inhibitors to decrease renin proteolysis. These steps included an ammonium sulfate precipitation and a batch-chromatography on DEAE-cellulose. The major purification step was an affinity chromatography on Sepharose-hexamethylene-diaminopepstatin. The enzyme obtained was further purified by molecular sieving gel filtration and isoelectric focusing.
由于肾素在肾脏中的浓度极低（不到总蛋白的1/50,000），其完全纯化面临着难题。从300千克新鲜猪肾开始，已大规模实现了猪肾肾素的完全纯化。获得了14.6毫克纯肾素，总产率为4%。纯化过程包括14个步骤。在pH 3.5下提取酶。后续纯化步骤在蛋白酶抑制剂存在下进行，以减少肾素的蛋白水解。这些步骤包括硫酸铵沉淀和DEAE - 纤维素柱层析。主要的纯化步骤是在琼脂糖 - 六亚甲基 - 二氨基胃蛋白酶抑制剂上进行亲和层析。所获得的酶通过分子筛凝胶过滤和等电聚焦进一步纯化。
