[Objective] The study was to determine the optimal fermentation process for Bacillus subtilis producing hirudin. [Method] With B. subtilis as the tested strain, the effects of activation time of strain, inoculation quantity, medium volume, temperature, initial pH value, carbon resource and nitrogen resource on fermentation were studied. And on the basis of that, the fermentation condition was optimized. [Result] The optimal fermentation condition was that temperature of 37 ℃, initial pH value of 7, medium volume of 20 ml /250 ml, activation time of strain of 6 h, inoculation quantity of 3%, nitrogen resource of 1.5% Tryptone, carbon resource of 9.0% soybean leaching solution, rotation rate of 220 r/min, fermentation time of 29 h and 0.3%Tween 80 was added after 16 h of fermentation. [Conclusion] Under the optimal condition, the activity of fermentation liquid could reach 210 ATU/ml, increasing 13.1 times than that before optimization. When the optimal fermentation condition was used in 50 L enlarged tank, the repeatability of test results was good, and the activity of fermentation liquid could reach 208 ATU/ml.%[目的] 确定芽孢杆菌发酵生产水蛭素的最佳工艺条件.[方法] 以枯草芽孢杆菌为供试菌种,研究菌种活化时间、接种量、培养基装量、温度、初始pH值、碳源、氮源等对发酵的影响,在此基础上对发酵条件进行优化.[结果] 最佳发酵条件为:温度37 ℃,初始pH值7,培养基装量20 ml/250 ml,菌种活化时间6 h,接种量3%,氮源为1.5%Tryptone,碳源为9.0%大豆浸出液,转速220 r/min,16 h后添加0.3%Tween 80,发酵时间29 h.[结论] 优化条件下,发酵液活性可达210 ATU/ml,较优化前提高13.1倍;优化条件上50 L罐放大后具有很好的重复性,发酵液活性可达208 ATU/ml.
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