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丙酸杆菌代谢物摇瓶补料分批发酵条件研究
孙 帅1, 常忠义1, 唐学明2
1.华东师范大学 生命科学学院;2.上海市农业科学院 生物技术研究所
摘要:
【目的】初步研究1株薛氏丙酸杆菌(Propionibacterium.shermanii)代谢物在摇瓶补料分批发酵中的最适碳源、氮源及其用量,旨在为丙酸杆菌工业化生产提供参考。【方法】在摇瓶分批发酵条件下,以丙酸杆菌对恶臭假单胞菌(Pseudomonas.pudia)的抑菌活性为检测指标,以未补加碳氮源处理为对照,从发酵第4~7天,每隔24 h补加1次不同质量浓度的复合碳源(m(乳酸钠)∶m(葡萄糖)=3∶1)、不同氮源(胰蛋白胨、酵母粉、牛肉膏),每天取样检测发酵液的抑菌活性,最后对获得最佳补料条件进行验证,在此基础上确定补料时间。【结果】从发酵第4天开始每隔24 h补加1次3 g/L复合碳源和2 g/L酵母粉时,丙酸杆菌代谢物对恶臭假单胞菌抑菌活性在第8 d最高,为33.47 AU/mL,显著高于对照组22.48 AU/mL(P<0.05);每隔24 h补料1次与每隔12 h补料1次,对提高抑菌活性的影响不大。【结论】在分批发酵过程中每隔24 h补加1次3 g/L复合碳源、2 g/L酵母粉,可以明显提高丙酸杆菌代谢物对恶臭假单胞菌的抑菌活性。
关键词:  薛氏丙酸杆菌  摇瓶补料分批发酵  抑菌活性
DOI:
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基金项目:上海市农业遗传育种重点实验室开放基金项目(Shagb2009-02);国家自然科学基金项目(30700064)
Study of Propionibacterium metabolites flask fed batch fermentation
Abstract:
【Objective】 The preliminary study was conducted to identify the optimal carbon and nitrogen sources and their dosage for Propionibacterium.shermanii under shake flask fed-batch fermentation conditions to provide reference for its industrial production.【Method】 Different mass concentrations of compound carbon source (m(lactate)∶m(glucose)=3∶1),various nitrogen sources (tryptone,yeast extract,beef extract) were added once every 24 h since the fourth day of the famentation in shake flask fed-batch fermentation conditions,using the antibacterial activity of the metabolites of Propionibacterium.shermanii against P.pudia as testing index,no additional carbon and nitrogen sources as CK.The antibacterial activity of fermentation broth was detected everyday.Finally the best feeding conditions and the feeding time were verified.【Result】 The results showed that when 3 g/L compound carbon source and 2 g/L yeast extract were added every 24 h since the fourth day of the famentation,the antibacterial activity of the metabolites of Propionibacterium.shermanii against P.pudia reached its highest antibacterial activity 33.47 AU/mL on the eighth day,significantly higher than that of the control group,22.48 AU/ml (P<0.05).But compared with feeding once every 24 h,feeding once every 12 h had little effect on the elevation of antibacterial activity.【Conclusion】 In shake flask fed-batch fermentation conditions,feeding 3 g/L compound carbon source and 2 g/L yeast extract once every 24 h can significantly improve the antibacterial activity of the metabolites of Propionibacterium.shermanii against P.pudia.
Key words:  Propionibacterium.shermanii  flask fed batch fermentation  antibacterial activity