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.
Vaz
F M
FM
Department of Clinical Chemistry and Pediatrics, University of Amsterdam, The Netherlands.
van Gool
S
S
Ofman
R
R
IJlst
L
L
Wanders
R J
RJ
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
D017886:Durapatite; D006899:Mixed Function Oxygenases; D050563:gamma-Butyrobetaine Dioxygenase; D002331:Carnitine