Purification and properties of alkaline phosphatase from porcine kidney / 医学研究生学报

ABSTRACT

Objective: For preparing the reference material of alkaline phosphatase(ALP), the purification and properties of ALP from porcine kidney were studied. Methods: The ALP purification procedure included isolation microvillus of porcine kidney cortices by solubilization the membrance-binding enzymes with 1-butanol, precipitation with ammonium sulfate, chromatography on DEAE-Sephacel,ConA-Sepharose,Sephadex G-200 and a immuno-affinity column of anti-GGT antibodies. Results: The purified enzyme had a specific activity of 402 kU/g protein at 37℃and was almost free of contaminating enzymes. The apparent Michaelis constants was 1.35 mmol/L, and the optimum pH was 10.40. Conclusion: The kinetic properties of the preparation were very close to those of the enzyme present in the human serum, The product was suitable as enzyme preparation for producing the enzyme reference material.