Carnitine biosynthesis. Purification of gamma-butyrobetaine hydroxylase from rat liver.
gamma-Butyrobetaine hydroxylase catalyse the last step in carnitine biosynthesis, the formation of L-carnitine from gamma-butyrobetaine, a reaction dependent on Fe2+, alpha-ketoglutarate, ascorbate and oxygen. Initial attempts to purify the protein from rat liver showed that gamma-butyrobetaine hydroxylase is unstable. We, therefore, determined the influence of various compounds on the stability of gamma-butyrobetaine hydroxylase at different storage temperatures. The enzyme activity was best conserved by storing the protein at 4 degrees C in the presence of 200 g/l glycerol and 10 mM DTT. We subsequently purified the enzyme from rat liver to apparent homogeneity by liquid chromatography.
T10 T11 T12 T13 T14 T15 T16 T17 T18 T1 T2 T3 T4 T5 T6 T7 T8 T9
CHEMICAL CHEMICAL CHEMICAL CHEMICAL CHEMICAL GENE-Y GENE-Y GENE-Y GENE-Y CHEMICAL CHEMICAL CHEMICAL CHEMICAL CHEMICAL CHEMICAL CHEMICAL CHEMICAL CHEMICAL
carnitine DTT L-carnitine Carnitine gamma-butyrobetaine gamma-Butyrobetaine hydroxylase gamma-butyrobetaine hydroxylase gamma-butyrobetaine hydroxylase gamma-butyrobetaine hydroxylase gamma-Butyrobetaine gamma-butyrobetaine Fe2+ alpha-ketoglutarate ascorbate oxygen gamma-butyrobetaine gamma-butyrobetaine glycerol