海豚链球菌诱变发酵法制备透明质酸及其在动物皮肤修复中的应用

Hyaluronic acid production by Streptococcus iniae and its application in rabbit skin’s regeneration

摘要:

透明质酸 (HA) 是一种非常重要的生物材料，是体内广泛存在的细胞外基质成分之一。为了获得产量、分子量及纯度较高的透明质酸，并研究透明质酸水凝胶在动物皮肤修复中的潜在作用。通过紫外诱变的方法对海豚链球菌进行诱变，并对此突变菌发酵后产物的蛋白含量及HA分子量进行了测定，通过CTAB法对发酵产物进行提纯，运用物理冻融法将透明质酸制成水凝胶后，用于兔背部全层皮肤修复的初探。结果表明通过诱变海豚链球菌产透明质酸的能力从 (82.3±3.3) mg/L增加到 (120±10.6) mg/L，增加了46.4%；产物经纯化后蛋白含量从 (0.178±0.011) mg/L减少到 (0.032±0.017) mg/L，减少了82.02%；所制得透明质酸的分子量约为3.0×105 Da；透明质酸水凝胶对兔全层皮肤缺损的修复有较明显的促进作用，能减轻炎症和伤口瘢痕的形成。

Abstract:

Hyaluronic acid (HA) is an important biomaterial as the extracellular matrix in human body. We produced HA by fermentation of Streptococcus iniae (Strep.). Production of HA by Strep. was evaluated and further improved by strain mutation by ultraviolet. One strain with higher HA yield and lower content of protein was obtained. Its HA yield increased from (82.3±3.3) mg/L to (120±10.6) mg/L, and protein decreased from (0.178±0.011) mg/L to (0.032±0.017) mg/L. The molecular weight (MW) of HA yield from Strep. is about 3.0×105 Da. Using the method of freezing and thawing, HA aqueous solution was transferred into hydrogel. This HA hydrogel, casted on sterilized non-woven fabric, was applied to repair rabbit skin with full-thickness defect. The preliminary results of the animal tests displayed that HA hydrogel obviously reduced the inflammation around the wound and promoted the skin regeneration comparing with the control tests.