y-Butyrobetaine hydroxylase catalyses the last step in carnitine biosynthesis, the formation of L-carnitine from y-butyrobetaine, a reaction dependent on Fe2+, a-ketoglutarate, ascorbate and oxygen. Initial attempts to purify the protein from rat liver showed that y-butyrobetaine hydroxylase is unstable. We, therefore, determined the influence of various compounds on the stability of y-butyrobetaine hydroxylase at different storage temperatures. The enzyme activity was best conserved by storing the protein at 4°C in the presence of 200 g/1 glycerol and l0mM DTT. We subsequently purified the enzyme from rat liver to apparent homogeneity by liquid chromatography.

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