枯草芽孢杆菌M364高产抗菌肽Bacillomycin D工业培养基优化

为降低抗菌肽工业发酵的生产成本,提高抗菌肽Bacillomycin D产量,在单因素基础上采用Plackett-Burman实验,寻找原始培养基6010中对Bacillomycin D产量影响显著的营养因素,并通过Central Composite Design响应面法对发酵培养基配方进行优化。结果表明:影响最显著的3个因素为:麦芽糖浆、尿素、MgSO₄·7H₂O。最佳工业发酵培养基配方为:麦芽糖浆46 g/L,尿素0.72 g/L,MgSO₄·7H₂O 0.35 g/L,玉米浆16 g/L,(NH₄)₂SO₄3 g/L,K₂HPO₄ 7 g/L,MnSO₄·H₂O 0.05 g/L。优化后,枯草芽孢杆菌M364摇瓶发酵Bacillomycin D产量达到(620.2±13.9)mg/L,比优化前(415.5±9.8)mg/L提高了50%,是常用Landy发酵培养基(240±20.6)mg/L的2.6倍;19 L发酵罐Bacillomycin D产量为(890.6±20.1)mg/L,相比优化后摇瓶的产量提高了44%,较原始6010培养基提高了1.14倍。本研究提供的培养基可同时达到高产,降低成本,缩短发酵时间的效果。

Optimization of Bacillomycin D High-yield Industrial Fermentation Medium of Bacillus subtilis M364

In order to reduce the cost of antibacterial peptide production in fermentation industry and to improve the yield of antibacterial peptide Bacillomycin D,Plackett-Burman design was utilized to evaluate the significant nutritional factors which affect the yield of Bacillomycin D in the original medium 6010 on the basis of the single factor experiment. And the industrial culture medium was optimized by the Central Composite Design response surface method. The results showed that the most three significant factors were maltose syrup,urea and MgSO₄·7H₂O. The optimum production was obtained when the cells grown in media containing:Maltose syrup 46 g/L,urea 0.72 g/L,MgSO₄·7H₂O 0.35 g/L,corn steep liquor 16 mL/L,(NH₄)₂SO₄ 3 g/L,K₂HPO₄ 7 g/L,MnSO₄·H₂O 0.05 g/L. After optimization,the yield of Bacillomycin D fermented by Bacillus subtilis M364 reached(620.2±13.9) mg/L,which increased by 50% compared to the non-optimized medium(415.5±9.8) mg/L and 2.6 times compared to the Landy fermentation medium(240±20.6) mg/L. The yield of Bacillomycin D in fermenter was(890.6±20.1) mg/L,which increased by 44% compared with the optimized production under the shaking flask condition and 1.14 times compared to the original 6010 medium. The culture medium provided by this study could achieve high yield,cost down and shorten fermentation time.