体外酶法水解制备小分子透明质酸

Preparation of Small Molecular Hyaluronan by Enzymatic Hydrolysis in Vitro

为了制备分子量分布集中的小分子透明质酸,在6 g/L的高分子量透明质酸水溶液中加入一定梯度重组水蛭透明质酸酶,并控制水解时间。结果表明,通过控制加入重组水蛭HAase的量和水解时间,根据水解分子量下降规律,可以制备出分子量分布集中的小分子HA,包括水解终产物透明质酸四糖和六糖。同时,通过高效体积排阻色谱和琼脂糖凝胶电泳,可以证明水解产物的分子量分布集中。

In order to get concentrated distribution of small molecular hyaluronan, 6 g/L of high molecu- lar hyaluronic acid aqueous solution was prepared, added with a certain gradient of hyaluronidase, and controlling the hydrolysis time. The results showed that controlling the amount of hyaluronidase and hy- drolysis time, particular molecular weight hyaluronic acid could be prepared according to the law of the hyaluronan molecular weight decreased, which included tetrasaccharide and hexasaccharide. Further- more,through high performance size exclusion chromatography and agarose gel electrophoresis, we could prove that the molecular weight distribution of hydrolysate was concentrated.