New enzymatic endpoint assay of serum sialic acid

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Assay of sialic acid in serum is useful for the diagnosis of inflammation, cancer, and other diseases [l-4]. The assay has been carried out chemically [5,6] and enzymatically. The enzymatic assay employing the reactions of neuraminidase (EC 3.2.1.18) and N-acetylneuraminic acid (NANA)-aldolase (EC 4.1.3.3) is composed of two groups. The one with lactate dehydrogenase (LDH, EC 1.1.1.27) is a UV-method [7], and the other with pyruvate oxidase (EC 1.2.3.3) and peroxidase (EC 1.11.1.7) is a calorimetric method [8]. These enzymatic assays are specific as well as simple. Recently, in our laboratory, N-acylmannosamine dehydrogenase (AMDH) was purified from a newly isolated bacterial strain (91. The enzyme oxidizes N-acylmannosamine in the presence of NAD+. It seems to be convenient for the assay of serum sialic acid. In this report, an enzymatic method for the estimation of sialic acid has been investigated using following reactions:

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