【摘要】 【目的】利用枯草芽胞杆菌(Bacillus subtilis SBS)进行联产血栓溶解酶和γ-聚谷氨酸研究【方法】本研究以实验室自行分离的Bacillus subtilis SBS为出发菌株,进行了液体发酵,通过正交实验研究了碳、氮源对血栓溶解酶和γ-聚谷氨酸联产的影响,并运用多种检测方法对产物进行了鉴定。【结果】在未添加谷氨酸的培养基中合成了γ-聚谷氨酸,表明该菌是非谷氨酸依赖型菌。合成血栓溶解酶的合适碳、氮源分别是可溶性淀粉和大豆蛋白胨,合成γ-聚谷氨酸的合适碳、氮源分别是蔗糖和NH4Cl。【结论】以蔗糖和大豆蛋白胨、NH4Cl分别作为碳源和氮源进行血栓溶解酶和γ-聚谷氨酸的联产。在蔗糖10 g/L、大豆蛋白胨20 g/L、NH4Cl 8 g/L时,血栓溶解酶酶活为265±25 IU/mL,γ-聚谷氨酸产量为1.183±0.015 g/L,均接近了单独合成时的水平。更多还原

【Abstract】 [Objective] To analyze the co-production of thrombolytic enzyme and γ-polyglutamic acid by liquid-culture of Bacillus subtilis SBS.[Methods] We used Bacillus subtilis SBS to produce thrombolytic enzyme（BTE） and γ-polyglutamic acid（（γ-PGA）） by liquid fermentation.We conducted orthogonal experiments analysis between carbon source and nitrogen source.Then through Fibrin-SDS-PAGE,UV spectrum,Infra-Red spectrum and high performance liquid chromatography（HPLC）,we identified the production.[Results] We synthesized γ-PGA in the medium without the addition of L-glutamic acid.Bacillus subtilis SBS was a L-glutamic acid-independent bacterium.The suitable carbon and nitrogen sources for the synthesis of thrombolytic enzyme（BTE） were soluble starch and soybean peptone.However,the suitable carbon and nitrogen sources for the synthesis of γ-PGA were sucrose and NH₄Cl.[Conclusion] The yields of both products could close to the levels of the single synthesis when the concentrations of sucrose,soybean peptone and NH₄Cl were 10,20,8 g/L respectively,the activity of BTE reached （265±25 IU/mL,）and the concentration of γ-PGA reached 1.183±0.015 g/L.更多还原