氧化胁迫联合UV、DES诱变选育高产、高分子量透明质酸菌株

Oxidative stress and UV,DES compound mutation screening of high molecular weight and high- yield hyaluronic acid strain

本实验以马疫链球菌MF003为出发菌,通过H2O2氧化胁迫培养、紫外辐照(UV)和硫酸二乙酯(DES)对菌株进行复合诱变,获得一株高产、高分子量透明质酸酶缺陷菌株ZD-25。经摇瓶发酵培养,该菌株透明质酸产量达到1.61g/L,相对分子量达到2.13×106u,分别比原始菌株提高了120.5%,57.8%。经过多次传代后,菌株透明质酸产量及相对分子量具有较高的稳定性。经5L发酵罐初步发酵,突变株ZD-25的透明质酸产量达到4.26g/L,相对分子量达到2.90×106u,分别比原始菌株提高了166.3%,93.3%。

Streptococcus equines MF003 was used as an original strain to perform H2O2 oxidative stress culture, ultraviolet radiation (UV) and diethyl sulfate (DES) compound mutation. After screening, a hyaluronidase deficient strain ZD-25 with high molecular weight and high-yield of hyaluronic acid was achieved. Based on shake flask fermentation, the yield of hyaluronic acid reached to 1.61 g/L and relative molecular weight of 2.13×10^6 u. Compared to the original strain,the yield and relative molecular weight were increased by 120.5% and 57.8%, respectively.The production and relative molecular weight of hyaluronic acid maintained high stability after several generation of reproduction.Furthermore,the yield of hyaluronic acid in mutant ZD-25 reached to 4.26g/L and relative molecular weight of 2.90×10^6u by 5L fermentation tank initial fermentation. Compared to the original strain, the yield and relative molecular weight increased 166.3% and 93,3%, respectively.