High-performance liquid chromatographic determination of four kinds of biotin-containing carboxylase.
A convenient and reliable reversed-phase liquid chromatographic assay for four kinds of carboxylase, i.e. pyruvate carboxylase, acetyl-coenzyme A carboxylase, propionyl-coenzyme A carboxylase, and 3-methylcrotonyl-coenzyme A carboxylase, was developed. These are all biotin enzymes, which contain covalently bound biotin as a co-factor (co-enzyme). This chromatographic method, which is free from interferences, gives a stoichiometric and precise assay for these enzymes. It is non-radioisotopic, which makes it suitable for clinical laboratories. The assay time required for one sample is ca. 30 min. The method was satisfactorily applied to guinea-pig kidney homogenates after a simple pretreatment with acid.
Oizumi
J
J
National Children's Medical Research Center, Division of Metabolism, Tokyo, Japan.
Hayakawa
K
K
D000103:Acetyl-CoA Carboxylase / Q000032:analysis*; D000818:Animals; D001710:Biotin / Q000032:analysis; D019735:Carbon-Carbon Ligases; D002262:Carboxy-Lyases / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006168:Guinea Pigs; D007668:Kidney / Q000201:enzymology; D008025:Ligases / Q000032:analysis*; D043586:Methylmalonyl-CoA Decarboxylase; D011766:Pyruvate Carboxylase / Q000032:analysis*
D001710:Biotin; D002262:Carboxy-Lyases; D008025:Ligases; D019735:Carbon-Carbon Ligases; D011766:Pyruvate Carboxylase; D000103:Acetyl-CoA Carboxylase; C021573:methylcrotonoyl-CoA carboxylase; D043586:Methylmalonyl-CoA Decarboxylase