食品科学 ›› 2017, Vol. 38 ›› Issue (8): 56-62.doi: 10.7506/spkx1002-6630-201708010

• 生物工程 • 上一篇    下一篇

丁酸梭菌的鉴定与发酵培养基配方优化

夏会丽,陈思思,陈 雄,代 俊,黄亚男,谢 婷,李爱玲,王 志   

  1. 1.湖北工业大学生物工程与食品学院,发酵工程教育部重点实验室,工业发酵湖北省协同创新中心, 湖北省工业微生物重点实验室,湖北 武汉 430068;2.河南省南街村(集团)有限公司,河南 临颍 462600; 3.天方药业股份有限公司,河南 驻马店 463000
  • 出版日期:2017-04-25 发布日期:2017-04-24
  • 基金资助:
    湖北省自然科学基金项目(2014CFB604)

Identification of Clostridium butyricum and Optimization of Fermentation Medium for Its Growth

XIA Huili, CHEN Sisi, CHEN Xiong, DAI Jun, HUANG Yanan, XIE Ting, LI Ailing, WANG Zhi   

  1. 1. Key Laboratory of Fermentation Engineering, Ministry of Education, Hubei Collaborative Innovation Center for Industrial Fermentation, Hubei Key Laboratory of Industrial Microbiology, School of Food and Biological Engineering, Hubei University of Technology, Wuhan 430068, China; 2. Nanjiecun (Group) Co. Ltd., Linying 462600, China; 3. Topfond Pharmaceutical Co. Ltd., Zhumadian 463000, China
  • Online:2017-04-25 Published:2017-04-24

摘要: 从河畔污泥中筛选出一株丁酸梭状芽孢杆菌(Clostridium butyricum HBUT-01),其16S rDNA核苷酸序列与Clostridium butyricum DSM 10702T菌的16S rDNA(AQQF01000149)的核苷酸序列同源性为99%。采用响应面法对丁酸梭菌HBUT-01的培养基配方进行优化,建立酵母粉、CaCO3和葡萄糖用量的二次回归模型,确定培养基最佳配方与质量分数为:葡萄糖1.60%、酵母粉0.72%、CaCO3 0.43%、蛋白胨2.50%、K2HPO4 0.05%、MgSO4·7H2O 0.08%、MnSO4·H2O 0.002%。10 L罐分批发酵实验确定了丁酸的比生成速率(qp)、细胞得率系数(Yx/s)分别比优化前降低了20%和提高了132%,说明优化后酸胁迫效应得到了缓解,细胞活性得到了有效的维持,使得生物量(14 h)达到了9.32×108 CFU/mL,是优化前(4.16×108 CFU/mL)的2.24 倍。

关键词: 丁酸梭菌, 16S rDNA, 培养基优化, 酸胁迫

Abstract: An anaerobic Clostridium strain designated HBUT-01was screened from river sludge. By 16S rDNA sequence analysis, the strain was identified as Clostridium butyricum and its 16S rDNA fragment shared 99% similarity with that of C. butyricum DSM 10702T (AQQF01000149). The response surface methodology (RSM) was used to optimize the fermentation medium for this isolated strain, and a quadratic regression model with yeast exact, CaCO3 and glucose as independent variables was established by Design-Expert 8.0 software. The optimum medium was determined as follows: glucose 1.60%, yeast extract 0.72%, peptone 2.50%, CaCO3 0.43%, K2HPO4 0.05%, MgSO4·7H2O 0.08%, and MnSO4·H2O 0.002%. Batch cultures were carried out in a 10-L bioreactor, and an increase in specific productivity of butyrate (qp) of 20% and an increase in biomass yield coefficient (Yx/s) of 132% were obtained as compared with that before optimization. The results suggested that the impact of acid stress on the cells was relieved effectively and cell activity was also maintained with the optimized medium, which resulted in an increased biomass, reaching 9.32×108 CFU/mL at 14 h, which was increased by 2.24 times as compared to that with the basic medium.

Key words: Clostridium butyricum, 16S rDNA, medium optimization, acid stress

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