Alkaline Phosphatases,Bacterial Alkaline,Calf Intestine,Molecular Cloning Experiments,MetalloEnzymes,Enzymatic Activity

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	Alkaline Phosphatases - Phosphatases are a group of enzymes which remove a phosphate from a variety of substrates like DNA, RNA and proteins. Phosphatases which acts in basic buffers with pH 8 or 9 are called as alkaline phosphatases. Most commonly bacterial alkaline phosphatases (BAP), calf intestine alkaline phosphatases (CIAP) and shrimp alkaline phosphatases are used in molecular cloning experiments.
The PO 4 from the substrate is removed by forming phosphorylated serine intermediate Figure 16.5. Alkaline phosphatase are metalloenzymes, and have Zn++ ions in them. BP A (bacterial alkaline phosphatase) is a dimer containing six Zn++ ions, two of which are essential for enzymatic activity. BP A is very stable and is not inactivated by heat and detergent.
Calf intestine alkaline phosphatases (CIAP) is also a dimer. It requires Zn++ and Mg++ ions for action. CIAP is inactivated by heating at 70°C for twenty min. or in the presence of 10 mM EGT A. Alkaline phosphatases are used to remove the PO 4 from the DNA or as reporter enzymes.