Pan Shihan, Chen Guiguang, Wu Rui, Cao Xiaoyan, Liang Zhiqun
State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi Microorganism and Enzyme Research Center of Engineering Technology, College of Life Science and Technology, Guangxi University, Nanning, China.
Front Microbiol. 2019 May 17;10:1025. doi: 10.3389/fmicb.2019.01025. eCollection 2019.
Microbial fibrinolytic enzyme is a promising candidate for thrombolytic therapy. Non-sterile production of fibrinolytic enzyme by marine D21-8 under submerged fermentation was realized at a mild temperature of 34°C, using a unique combination of starvation strategy and self-production of antibacterial agents. A medium composed of 18.5 g/L glucose, 6.3 g/L yeast extract, 7.9 g/L tryptone, and 5 g/L NaCl was achieved by conventional and statistical methods. Results showed efficient synthesis of fibrinolytic enzyme and antibacterial compounds required the presence of both yeast extract and tryptone in the medium. At shake-flask level, the non-sterile optimized medium resulted in higher productivity of fibrinolytic enzyme than the sterile one, with an enhanced yield of 3,129 U/mL and a production cost reduced by 24%. This is the first report dealing with non-sterile submerged fermentation of fibrinolytic enzyme, which may facilitate the development of feasible techniques for non-sterile production of raw materials for the preparation of potential drugs with low operation cost.
微生物纤溶酶是溶栓治疗的一个有前景的候选药物。利用饥饿策略和自身产生抗菌剂的独特组合，在34°C的温和温度下，通过海洋D21-8在深层发酵中实现了纤溶酶的非无菌生产。通过常规方法和统计方法获得了一种由18.5 g/L葡萄糖、6.3 g/L酵母提取物、7.9 g/L胰蛋白胨和5 g/L氯化钠组成的培养基。结果表明，高效合成纤溶酶和抗菌化合物需要培养基中同时存在酵母提取物和胰蛋白胨。在摇瓶水平上，非无菌优化培养基产生的纤溶酶生产率高于无菌培养基，产量提高到3129 U/mL，生产成本降低了24%。这是第一篇关于纤溶酶非无菌深层发酵的报道，这可能有助于开发可行的技术，以低成本非无菌生产制备潜在药物的原料。
