Purification tissue storage

Of all the enzymes of the carnitine biosynthetic pathway, yBBH is the best-studied enzyme. Like e-N-trimethyllysine hydroxylase, y-BBH is a non-heme ferrous-iron dioxygenase that requires a-ketoglutarate, Fe and molecular oxygen as cofactors. In this class of enzymes, the hydroxylation of the substrate is linked to the oxidative decarboxylation of a-ketoglutarate. Ascorbate is needed to maintain iron in die reduced state. yBBHhas been isolated from various sources including human kidney, " catf and rat liver and the bacterium Pseudomonas AKl. A common problem in the purification of this enzyme from mammalian tissues is the poor stability of the protein. We, therefore, determined the optimal storage conditions and subsequently pmified ybutyrobetaine hydroxylase from rat liver. 

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