Purification and properties of alkaline phosphatase from the mucosa of rat small intestine

Alkaline phosphatase [orthophosphoric monoester phosphohydrolase, EC 3.1.3.1] was purified from the mucosa of rat small intestine by butanol extraction, ethanol fractionation, gel filtration, with controlled-pore glass-10 and DEAE-cellulose column chromatography. On the gel filtration, the enzyme ac...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 1979-11, Vol.86 (5), p.1225-1231
Main Authors: Nakasaki, H, Matsushima, T. (Tokyo Univ. (Japan). Inst. of Medical Science), Sato, S, Kawachi, T
Format: Article
Language:English
Subjects:
Alkaline Phosphatase - isolation & purification
Alkaline Phosphatase - metabolism
Animals
Drug Stability
Hot Temperature
Intestinal Mucosa - enzymology
Intestine, Small - enzymology
Kinetics
Leucine - pharmacology
Male
Molecular Weight
Phenylalanine - pharmacology
Rats
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Summary:Alkaline phosphatase [orthophosphoric monoester phosphohydrolase, EC 3.1.3.1] was purified from the mucosa of rat small intestine by butanol extraction, ethanol fractionation, gel filtration, with controlled-pore glass-10 and DEAE-cellulose column chromatography. On the gel filtration, the enzyme activity was separated into three peaks; A in the void volume, B and C at lower molecular weight positions. Enzyme A was purified to homogeneity. The activity of enzymes A, B, and C was detected even on sodium dodecyl sulfate-polyacrylamide gel electrophoresis at the position of the protein of enzyme A, which had a molecular weight of 110,000 daltons. Enzymatic properties such as pH optimum, Km value for the substrate, heat inactivation and inhibition by amino acids were the same in all three enzymes. Based on these findings, together with the elution positions on gel filtration, enzyme A was regarded as an aggregate, and enzymes B and C as dimer and monomer molecules, respectively.
ISSN:0021-924X
1756-2651
DOI:10.1093/oxfordjournals.jbchem.a132637