Large scale preparation of pure hog kidney renin.

ABSTRACT

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.