采用单因素试验和正交试验对具有水质净化功能的解淀粉芽孢杆菌HN的培养基成分及培养条件进行了优化,并采用100 L发酵罐对优化发酵技术参数进行了验证试验。结果表明,优化的培养基成分为:蛋白胨2.0g/L,牛肉膏3.0g/L,可溶性淀粉7.0g/L,氯化钠6.0g/L|培养条件为:培养基的初始pH值为7.0,装液量为100mL/250mL,接种量为3%,温度为37℃,发酵周期24～30 h|在上述优化培养条件下的生长曲线结果显示,菌株HN在9 h后进入对数生长期,24 h达到稳定期,菌液浓度达35.6×108cfu/mL,最高芽孢浓度达到30.4×108cfu/mL|100 L发酵罐验证试验表明,37℃发酵26 h活菌浓度可达37.0×108cfu/mL,芽孢浓度为33.6×108cfu/mL。
The single factor test and orthogonal-design experiment were applied to screen optimal fermentation technology such as medium composition and fermentation conditions for Bacillus amyloliquefaciens HN with water purification,and the fermentation parameters in fermentor of 100 L was scaled up successfully. The results showed that the strain HN was easy to cultivate in a medium with 2.0g/L peptone,3.0g/L beef extract,7.0g/L soluble starch,6.0g/L NaCl and the optimum fermentation conditions were initial culture medium pH 7. 0,inoculation rate 3%(V/V),100mL/250mL medium volume at 37℃ for 24-30 h. Under this condition,strain HN reached loga-rithmic phase at 9 h,came to stable period after cultured at 37℃ for 24 h and the bacteria concentration reached 35. 6×108cfu/mL and the spore concentration was up to 30.4×108cfu/mL in 250mL triangular flask. The valida-tion test in 100 L fermentor result indicated that bacteria number and spore concentration reached 37. 0×108 cfu/mL and 33. 6×108cfu/mL,respectively after fermented at 37℃ for 26 h.
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