海洋侧孢短芽孢杆菌S-12-86溶菌酶的分离纯化与冻干技术研究

Purification and lyophillization of Lysozyme produced by marine microorganism

中文关键词: 海洋微生物  溶菌酶  分离纯化；冷冻干燥

英文关键词: Marine microorganism  Lysozyme  Purification  Lyophillization

中文摘要:

      海洋微生物溶菌酶发酵液经低温离心、超滤浓缩、乙醇提取、Superdex 75 10/300凝胶层析和反相高效液相色谱层析纯化得到电泳纯海洋溶菌酶，分子量为17.1 kD，比活达到3987.7 U/mg，纯度提高41.98倍，活力回收率为21.7%。对该酶冷冻干燥过程的研究结果表明，海藻糖、蔗糖和麦芽糖对该酶均有一定的冻干保护作用。其中，以海藻糖的保护效果最佳。0.5 mol/L海藻糖与20 mg/ml吐温80复合作为冻干保护剂，使冻干后的溶菌酶活性维持在95%以上，为该酶进一步研究和应用提供了稳定的酶制剂。

英文摘要:

      Cell-free supernatant with marine microorganism lysozyme was prepared by centrifugation of culture broth，ultrafiltration and concentration. The crude lysozyme was purified 41.98 fold to electrophoretic homogeneity with recovery of 21.7％ and specific activity of 3 987.7 U/mg by extracting with ethanol, Superdex 75 10/300 chromatography and reversed-phase HPLC. The relative molecular weight of this lysozyme was 17.1 kD determined by SDS-PAGE electrophoresis. Preparations of lysozyme from native marine microorganism were formulated with different additives for lyophillization. The studied additives, including trehalose, sucrose and maltose, showed good protecting effect with trehalose showing the best performance for freeze-drying stabilization. Comparing with the native lysozyme in the absence of protective agents, the activity of the freeze-dried lysozyme treated by trehalose (0.5 mol/L) and Tween 80 (20 mg/L) as protective agents retained more than 95% of the original activity.