The action of Streptomyces hyalurolyticus hyaluronate lyase on hyaluronic acid (HA) determined by high-performance capillary electrophoresis (HPCE) (electrokinetic chromatography with sodium dodecyl sulphate) was examined and compared with the HPLC procedure. By using an uncoated fused-silica capillary tube, 50 mum ID, 65 cm long from the injection point to the detector, the separation of HA species from DP 4 to approx. DP 30 was obtained. The length of the capillary was found to be fundamental for the separation and resolution of the unsaturated HA oligosaccharides. Furthermore, the HPCE separation of HA Delta-tetrasaccharide and Delta-hexasaccharide species produced a greater detection sensitivity (about 20 times greater) than HPLC. Various HA samples were analyzed for the ratio of tetrasaccharide/hexasaccharide (T/H) after treatment with S. hyalurolyticus hyaluronidase. HA samples of extractive origin of different molecular masses showed a T/H ratio between 1.28 and 1.48 determined by HPCE, with a good correspondence with the HPLC separation. On the contrary, a sample of cross-linked HA resulted in a great discrepancy between the two analytical techniques (greater than 40%). HA of fermentative origin had a T/H ratio of approx. 1.92-2.00, close to that of HA (approx. 1.80) for the first time extracted and purified from the body of a species of mollusc bivalve, Mytilus galloprovincialis. The presence of resistant (less susceptible to enzyme cleavage) site repeating unit in the carbohydrate backbone of HA is discussed in relation with the T/H values experimentally determined by HPCE.
|Anno di pubblicazione:||2004|
|Titolo:||High-performance capillary electrophoresis separation of hyaluronan oligosaccharides produced by Streptomyces hyalurolyticus hyaluronate lyase.|
|Autore/i:||F. Maccari; F. Tripodi; N. Volpi|
|Codice identificativo ISI:||WOS:000221364700009|
|Codice identificativo Scopus:||2-s2.0-2042542045|
|Citazione:||High-performance capillary electrophoresis separation of hyaluronan oligosaccharides produced by Streptomyces hyalurolyticus hyaluronate lyase. / F. Maccari; F. Tripodi; N. Volpi. - In: CARBOHYDRATE POLYMERS. - ISSN 0144-8617. - STAMPA. - 56(2004), pp. 55-63.|
|Tipologia||Articolo su rivista|
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