Transphosphorylation mechanism of hen's egg yolk alkaline phosphatase

1. Hen's egg yolk alkaline phosphatase can transfer the donor substrate phosphoryl to a hydroxyl containing acceptor. 2. This acceptor contains preferably an alpha-amino or alpha-imino group. 3. The optimal pH of the transfer reaction is at 9.45 with Tris and at 9.60 with diethanolamine as acceptor. 4. Transfer products were isolated and characterized. 5. The ratio of products is independent of the donor substrate. ROH and R'OP production increase linearly with the acceptor concentration, while Pi release and Km remain constant. 6. This points to a mechanism involving two phosphorylated intermediates. 7. The acceptor bypasses the isomerization step of the phosphoryl enzyme, which is necessary before hydrolysis can take place.