Carnitine biosynthesis. Purification of gamma-butyrobetaine hydroxylase from rat liver.

F M Vaz, S van Gool, R Ofman, L IJlst, R J Wanders

Department of Clinical Chemistry and Pediatrics, University of Amsterdam, The Netherlands.

Advances in experimental medicine and biology 1999

filter terms:

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.

Citation

F M Vaz, S van Gool, R Ofman, L IJlst, R J Wanders. Carnitine biosynthesis. Purification of gamma-butyrobetaine hydroxylase from rat liver. Advances in experimental medicine and biology. 1999;466:117-24