Fructose-1, 6-diphosphate Aldolase UV Assay, Manuel Method

Publisher Summary This chapter focuses on aldolase, which was first crystallized in 1943 by Warburg and Christian from rat muscle. The enzyme is widely distributed; it is found in all cells which catabolize carbohydrate via glycolysis. Fructose-1,6-diphosphate aldolase is applied in biochemistry and clinical chemistry. The pyruvate contained in the sample reacts with lactate dehydrogenase and NADH before the start of the assay. With collidine buffer the aldolase activity of serum, blood hemolysates, and muscle homogenates has a broad pH optimum between 7 and 8, while with veronal buffer it is between 8.5 and 9. In the presence of cyanide, crystalline aldolase from bovine liver has a pH optimum between 9.1 and 9.4 with fructose-1,6-diphosphate and between 8.1 and 8.4 with fructose-1-phosphate. All solutions should be prepared with fresh doubly distilled water. Store all solutions at ca 4°C. Aldolase is relatively thermostable in the presence of large amounts of other proteins. The crystalline enzyme obtained from ox liver catalyses the cleavage of fructose-1-phosphate and the synthesis of erythrulose phosphate from dihydroxyacetone phosphate to about the same extent, which suggest that existence of separate enzymes for these reactions is unlikely.