A neuraminidase from Streptococcus sanguis that can release O-acetylated sialic acids.
The naturally occurring sialic acids can have different types of N- and O-substitutions, resulting in more than 20 known isomers and compounds. Most methods for the detailed study of these various sialic acids require that the molecules be first released from their alpha-glycosidic linkage. When mild acid hydrolysis is used for this purpose, significant destruction of O-substituent groups occur. On the other hand, the presence of O-substituent groups renders the sialic acid molecule partially or completely resistant to the action of the currently known neuraminidase. To circumvent this problem, we searched for a neuraminidase whose activity is not affected by O-substitution. We reasoned that because Streptococcus sanguis from the human oral cavity is continually exposed to O-substituted sialic acids, its extracellular neuraminidase might not be blocked by O-substitution. We therefore purified this enzyme 3100-fold (56% yield) using ammonium sulfate precipitation, N-(p-aminophenyl)oxamic acid-agarose affinity chromatography, and chromatography on quaternary aminoethyl (QAE)-Sephadex, sulfopropyl (SP)-Sephadex, and Sephacryl S-200. The purified preparation is free of other significant glycosidase activities and proteolytic activities. It is capable of quantitatively releasing all the O-acetylated sialic acids that we studied with the single exception of the 4-O-acetylated sialic acid of equine submaxillary mucin. The activity of the enzyme is also not restricted by the type pf sialic acid linkage or the nature of the underlying oligosaccharide. However, it has maximal activity on gangliosides only in the presence of detergents. The general properties of this enzyme are described and its substrate specificities are contrasted with those of the commonly used neuraminidase from Vibrio cholerae.
D000818:Animals; D002417:Cattle; D007700:Kinetics; D009077:Mucins; D009439:Neuraminidase / Q000378:metabolism*; D012794:Sialic Acids / Q000302:isolation & purification / Q000378:metabolism*; D013298:Streptococcus sanguis / Q000201:enzymology*; D013363:Submandibular Gland; D013379:Substrate Specificity
D009077:Mucins; D012794:Sialic Acids; D009439:Neuraminidase