Blautia coccoidesGA-1的传代培养及其同化H2/CO2产乙酸特征

doi:10.11949/j.issn.0438-1157.20151884

化工学报 ›› 2016, Vol. 67 ›› Issue (6): 2591-2597.DOI: 10.11949/j.issn.0438-1157.20151884

Blautia coccoidesGA-1的传代培养及其同化H₂/CO₂产乙酸特征

刘崇¹, 李建政^1,2, 张亚非¹, 张玉鹏¹, 尤立剑³, 刘冬梅¹

1. 哈尔滨工业大学市政环境工程学院, 黑龙江哈尔滨 150090;
2 . 哈尔滨工业大学城市水资源与水环境国家重点实验室, 黑龙江哈尔滨 150090;
3. 黑龙江大庆市大同区环保监测站, 黑龙江大庆 163515

收稿日期:2015-12-11 修回日期:2016-01-22 出版日期:2016-06-05 发布日期:2016-06-05
通讯作者: 李建政
基金资助:
国家自然科学基金项目(51178136)。

Sub-culturing of novel Blautia coccoides GA-1 and acetate synthesis from H₂/CO₂ in subcultures

LIU Chong¹, LI Jianzheng^1,2, ZHANG Yafei¹, ZHANG Yupeng¹, YOU Lijian³, LIU Dongmei¹

1. School of Municipal and Environmental Engineering, Harbin Institute of Technology, Harbin 150090, Heilongjiang, China;
2. State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin 150090, Heilongjiang, China;
3. Environmental Monitoring Station of Datong District, Daqing 163515, Heilongjiang, China

Received:2015-12-11 Revised:2016-01-22 Online:2016-06-05 Published:2016-06-05
Supported by:
supported by the National Natural Science Foundation of China (51178136).

摘要/Abstract

摘要：

为了解同型产乙酸菌异养代谢与自养代谢的相互作用与机制,并为快速获得具有较强自养代谢能力的菌体细胞提供培养方法,以H₂/CO₂和(或)葡萄糖为碳源,考察了Blautia coccoidesGA-1在连续传代培养中的代谢特征。结果表明,以H₂/CO₂作为唯一碳源进行连续传代培养时,菌株GA-1长势较弱,其子代的自养代谢能力也逐渐下降;在葡萄糖培养基中,菌株GA-1增殖旺盛,但高浓度的葡萄糖对其自养代谢能力有显著抑制作用,这种抑制作用可能是自养代谢和异养代谢对辅酶A和ATP的竞争、酸性环境造成的代谢抑制以及辅酶I的氧化还原平衡调节等综合作用的结果。以体积比为4:1的H₂/CO₂混合气为气相条件,用200mg·L^-1葡萄糖培养基对菌株GA-1进行传代培养,不仅可获得稳定的子代培养物,而且可以将其利用H₂/CO₂产乙酸的能力维持在2.16g乙酸·(g干细胞)^-1的水平。

关键词: 同型产乙酸菌, 发酵, 代谢, 氢, 二氧化碳, 乙酸

Abstract:

Though homoacetogens have been defined as autotrophic anaerobes using acetyl-CoA Wood-Ljungdahl carbon fixation pathway (WLP) for energy conservation, acetate production and biomass formation from CO₂, these anaerobes can also utilize organic compounds heterotrophically to produce acetate and form biomass by Embden-Meyerhof-Parnas (EMP) glycolysis. A better understanding of the interrelationship between autotrophy and heterotrophy in homoacetogens will be helpful for acetate biosynthesis from exhaust rich in CO₂. However, interaction of the two metabolisms has not been well understood though both of which have been extensively investigated respectively. Blautia coccoides GA-1, a novel homoacetogen strain obtained in a previous study, was introduced to the present research and the interrelationship between autotrophy and heterotrophy in the strain was investigated by sub-culturing with glucose and (or) H₂/CO₂ (vol, 4:1) as carbon source (s). The results showed that the cell production of strain GA-1 in H₂/CO₂ cultures was too low to be sub-cultured after the 5th generation due to the energy produced was very limited in WLP. On the contrary, the strain could grow very well in glucose cultures and could be sub-cultured steadily. However, the autotrophic capability of the subcultures decreased by generations, even lost since the 7th generation in 1000 mg·L^-1 glucose cultures. This may be resulted in: (1) the competition for Coenzyme A (CoA) between the autotrophic WLP and the heterotrophic EMP glycolysis, (2) ATP produced in the strain used up for biosynthesis rather for acetate synthesis from H₂/CO₂, (3) inhibition of lower pH less than 5.5 to the activity of hydrogenase, (4) negative effect of the lower pH on intracellular balance of NADH/NAD⁺. In mixotrophic cultures with H₂/CO₂ and 200 mg·L^-1 glucose as co-carbon sources, strain GA-1 could not only be sub-cultured steadily but also keep autotrophic activity well in the generations with a specific acetate production of about 2.16 g·(g dry cell)^-1 from H₂/CO₂.

Key words: homoacetogens, fermentation, metabolism, hydrogen, carbon dioxide, acetate

中图分类号:

X172

刘崇, 李建政, 张亚非, 张玉鹏, 尤立剑, 刘冬梅. Blautia coccoidesGA-1的传代培养及其同化H₂/CO₂产乙酸特征[J]. 化工学报, 2016, 67(6): 2591-2597.

LIU Chong, LI Jianzheng, ZHANG Yafei, ZHANG Yupeng, YOU Lijian, LIU Dongmei. Sub-culturing of novel Blautia coccoides GA-1 and acetate synthesis from H₂/CO₂ in subcultures[J]. CIESC Journal, 2016, 67(6): 2591-2597.

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Blautia coccoidesGA-1的传代培养及其同化H₂/CO₂产乙酸特征

Sub-culturing of novel Blautia coccoides GA-1 and acetate synthesis from H₂/CO₂ in subcultures

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