ImmobilizedE. coli Alkaline Phosphatase |
| |
Authors: | Khaja Basheeruddin Vicki Rothman Simeon Margolis |
| |
Affiliation: | (1) Johns Hopkins University School of Medicine, 720 Rutland Avenue, 21205 Baltimore, Maryland |
| |
Abstract: | We have immobilized E.coli alkaline phosphatase (EC 3.1.3.1) by linking it covalently to sepharose 4B. This preparation has several advantages over the soluble enzyme. The immobilized enzyme is easily separable from other constituents in incubation mixtures. The immobilized enzyme can be reused repeatedly and is more stable than the soluble enzyme to heat treatment in the presence of 10 mM Mg2+. The insoluble and soluble phosphatases removed 75 and77%, respectively, of the inorganic phosphorus from casein. The immobilized enzyme inactivated two enzymes believed to be active in the phosphorylated state, acyl-CoA : cholesterol acyltransferase (ACAT) by 39% and NADPH-cytochrome P-450 reductase by 89%. The utility of immobilized alkaline phosphatase for studying the phosphorylation and dephosphorylation of soluble or membrane-bound enzymes and proteins is discussed. |
| |
Keywords: | Alkaline phosphatase, immobilized immobilized alkaline phosphatase, from E.coli stability, of immobilized alkaline phosphatase protein dephosphorylation casein Acyl-CoA: cholesterol acyltransferase dephosphorylation, immobilized alkaline phosphatase and |
本文献已被 SpringerLink 等数据库收录! |
|