Large scale purification of hog renin. Physicochemical characterization

Area of Science:

• Biochemistry
• Enzymology

Background:

• Renin is a key enzyme in the renin-angiotensin system, regulating blood pressure.
• Purification of active renin is essential for its comprehensive characterization.

Purpose of the Study:

• To develop a purification strategy for hog kidney renin.
• To perform enzymatic and physicochemical characterization of purified renin.

Main Methods:

• Extraction from hog kidney with protease inhibitors.
• Multiple chromatographic techniques including ion exchange and gel filtration.
• Isoelectric focusing and SDS-gel electrophoresis for purity assessment.

Main Results:

• Obtained 2.3 mg of renin with 70,000-fold purification and 16% recovery.
• Confirmed purity using SDS-PAGE and polyacrylamide gel electrophoresis.
• Characterized renin's stability, molecular weight (36,800 Da), isoelectric point (5.15), and Michaelis constant (Km = 7.7 x 10^-6 M).

Conclusions:

• Successfully purified hog kidney renin to homogeneity.
• Renin is a glycoprotein with defined physicochemical properties.
• The characterized properties provide a basis for further functional studies.