title: Carnitine biosynthesis. Purification of gamma-butyrobetaine hydroxylase from rat liver.
abstract: 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.
authors.affiliation: Department of Clinical Chemistry and Pediatrics, University of Amsterdam, The Netherlands.
authors.forename: F M
authors.initials: FM
authors.lastname: Vaz
authors.forename: S
authors.initials: S
authors.lastname: van Gool
authors.forename: R
authors.initials: R
authors.lastname: Ofman
authors.forename: L
authors.initials: L
authors.lastname: IJlst
authors.forename: R J
authors.initials: RJ
authors.lastname: Wanders
mesh_terms: D000818:Animals; D002331:Carnitine / Q000096:biosynthesis*; D002845:Chromatography; D002852:Chromatography, Ion Exchange; D017886:Durapatite; D004795:Enzyme Stability; D008099:Liver / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D051381:Rats; D050563:gamma-Butyrobetaine Dioxygenase
chemical_list: D017886:Durapatite; D006899:Mixed Function Oxygenases; D050563:gamma-Butyrobetaine Dioxygenase; D002331:Carnitine