食品科学 ›› 2023, Vol. 44 ›› Issue (10): 158-164.doi: 10.7506/spkx1002-6630-20220423-300

• 生物工程 • 上一篇    下一篇

酪丁酸梭菌共培养对速生梭菌己酸代谢的影响及机理探讨

陈茂彬,殷想想,郭志豪,晋湘宜,李良,董孝元,常煦,方尚玲   

  1. (1.湖北工业大学生物工程与食品学院,湖北 武汉 430068;2.黄鹤楼酒业有限公司,湖北 武汉 430050;3.湖北安琪酵母股份有限公司,湖北 宜昌 443000)
  • 出版日期:2023-05-25 发布日期:2023-06-02
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2016YFD0400500);湖北省科技厅重大专项(2018ABA084)

Effect and Mechanism of Co-culture with Clostridium tyrobutyricum on Caproic Acid Mechanism in Clostridium celerecrescens

CHEN Maobin, YIN Xiangxiang, GUO Zhihao, JIN Xiangyi, LI Liang, DONG Xiaoyuan, CHANG Xu, FANG Shangling   

  1. (1. School of Biological Engineering and Food, Hubei University of Technology, Wuhan 430068, China; 2. Yellow Crane Tower Distillery Co. Ltd., Wuhan 430050, China;3. Angel Yeast Co. Ltd., Yichang 443000, China)
  • Online:2023-05-25 Published:2023-06-02

摘要: 以1 株产己酸的速生梭菌(Clostridium celerecrescens)JSJ-1与1 株产丁酸的酪丁酸梭菌(Clostridium tyrobutyricum)D-1为研究对象,通过不同的培养方式(固体、液体、透析袋)比较两种微生物共培养与纯培养生长代谢的差异,通过在纯培养JSJ-1时添加不同质量浓度丁酸和丁酸钠,近一步探究了共培养时丁酸菌对己酸菌己酸代谢的影响及机理。结果表明:34 ℃时,D-1与JSJ-1同时接种于含葡萄糖的复合固体培养基厌氧培养,D-1比JSJ-1更具有生长优势,会优先利用葡萄糖进行生长。D-1与JSJ-1在液体培养基中培养,共培养己酸的生成时间比相同条件下JSJ-1单独培养时提前了3 d,第12天时己酸质量浓度分别为9.93 g/L和9.79 g/L。葡萄糖对JSJ-1生成己酸有较强的抑制作用,共培养时D-1优先利用葡萄糖可缓解葡萄糖对JSJ-1合成己酸的抑制作用。将JSJ-1与D-1通过直接液体培养与加透析袋共培养作对比,结果表明,JSJ-1与D-1共培养时,JSJ-1与D-1是否存在细胞间的直接接触对己酸菌己酸产量无影响。通过向纯培养己酸菌培养基中分别添加丁酸和丁酸钠,近一步得出D-1在pH 6~6.5时生成的丁酸能够促进JSJ-1合成己酸。共培养时,D-1优先利用葡萄糖不仅可以缓解葡萄糖对己酸生成的抑制作用,而且D-1生成的丁酸可以促进己酸的合成。

关键词: 速生梭菌;酪丁酸梭菌;共培养;纯培养

Abstract: This study compared the differences in the growth and metabolism of Clostridium celerecrescens JSJ-1, a strain able to produce caproic acid, and Clostridium tyrobutyricum D-1, a stain able to produce butyric acid, when cultured separately or co-cultured in different systems (solid medium, and liquid medium with or without use of dialysis bag). Furthermore, this study investigated the effect and mechanism of co-culture with C. tyrobutyricum on caproic acid metabolism in caproic acid-producing bacteria by culturing JSJ-1 alone in the presence of butyric acid or sodium butyrate at different concentrations. The results showed that when co-cultured on a composite solid medium containing glucose in anaerobic conditions at 34 ℃, D-1 grew better than JSJ-1, and could preferentially utilize glucose for growth. The generation time of caproic acid was three days earlier when D-1 and JSJ-1 were co-cultured in a liquid medium than when JSJ-1 was cultured alone under the same conditions, and the concentration of caproic acid was 9.93 and 9.79 g/L on the 12th day in the two cases, respectively. Glucose had a strong inhibitory effect on the production of caproic acid by JSJ-1, which could be relieved by preferential utilization of by D-1 upon co-culture. The co-culture in the liquid medium with and without use of dialysis bag were compared, revealing that whether there was direct contact between JSJ-1 and D-1 had no effect on the yield of caproic acid. By purely culturing JSJ-1 in the presence of butyric acid or sodium butyrate, it was found that butyric acid generated by D-1 at pH 6–6.5 could promote the synthesis of caproic acid by JSJ-1. During co-culture, the preferential utilization of glucose by D-1 could relieve the inhibitory effect of glucose on the production of caproic acid, and butyric acid produced by D-1 could promote the synthesis of caproic acid.

Key words: Clostridium celerecrescens; Clostridium tyrobutyricum; pure culture; co-culture

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