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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2014, Vol. 34 Issue (9): 48-55    DOI: 10.13523/j.cb.20140908
研究报告     
阻断嗜乙酰乙酸棒杆菌乙酸合成途径对其在缺氧条件下产琥珀酸的影响
刘伟, 郑璞, 靳新娜
江南大学生物工程学院 工业生物技术教育部重点实验室 无锡 214122
Effects of Disrupting Acetate Formation Pathways in Corynebacterium acetoacidophilum on Succinate Production Under Oxygen Deprivation
LIU Wei, ZHENG Pu, JIN Xin-na
The Key Laboratory of Industrial Biotechnology, Ministry of Education; School of Biotechnology, Jiangnan University, Wuxi 214122, China
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摘要：

为探究和阻断嗜乙酰乙酸棒杆菌乙酸代谢途径，提高缺氧条件下琥珀酸的产率，减少副产物乙酸的合成，以C. acetoacidophilum ΔldhA为出发菌株，利用同源重组的方法分别敲除磷酸乙酰转移酶、乙酸激酶、CoA转移酶和丙酮酸脱氢酶复合体的相关基因pta，ackA，ctfA与aceE，研究突变菌产琥珀酸过程中相关参数的变化。结果表明：敲除pta与ackA基因后，对乙酸浓度，糖耗速率和糖酸转化率影响不大；pta，ackA与ctfA基因的同时失活使得乙酸的浓度和摩尔转化率分别降低81.4%和77.2%，葡萄糖消耗速率下降28.3%，琥珀酸对葡萄糖摩尔转化率提高25.3%；而单独敲除aceE基因后，乙酸几乎不产生，葡萄糖消耗速率下降35.6%，琥珀酸对葡萄糖摩尔转化率提高34.7%。因此，缺氧条件下，嗜乙酰乙酸棒杆菌的乙酸合成几乎全部走乙酰CoA途径，pta，ackA与ctfA是由乙酰CoA合成乙酸途径中最主要的基因；敲除基因aceE， 可以完全阻断乙酸生成，有效提高琥珀酸产率。
关键词： 嗜乙酰乙酸棒杆菌;  乙酸;  琥珀酸;  代谢阻断    
Abstract:

In order to reduce acetate production and improve succinate production and explore the main acetate formation pathways of C.acetoacidophilum, gene of pta, ackA, ctfA and aceE, which encoded phosphotransacetylase, acetate kinase, CoA-transfease and pyruvate dehydrogenase complex in C.acetoacidophilum were disrupted respectively by the means of homologous recombination. In C.acetoacidophilum ΔldhAΔpta-ackA, the titer of acetate, gluocse consumption rate, and both yields of acetate and succinate were found to be similar to that of C.acetoacidophilum ΔldhA. Furthermore, the titer and yield of acetate decreased by 81.4% and 77.2%, respectively, the glucose consumption rate decreased by 28.3% and the yield of succinate increased by 25.3% in C.acetoacidophilum ΔldhAΔctfAΔpta-ackA. In addition, when gene aceE was deleted, C. acetoacidophilum ΔldhAΔaceE almost produced no acetate, the glucose consumption rate decrease by 35.6% and the yield of succinate increased by 34.7%. Under oxygen deprivation, almost all acetate formed through acetyl-CoA in C.acetoacidophilum. Gene pta, ackA and ctfA were the main genes of acetate formation pathways from acetyl-CoA. Disrupting gene aceE could cut off acetate synthesis in C.acetoacidophilum, and effectively increase succinate production.
Key words: Corynebacterium acetoacidophilum    Acetate    Succinate    Metabolic disruption
收稿日期: 2014-06-30 出版日期: 2014-09-25
ZTFLH:  Q784  
基金资助:

国家“863”计划（2006AA020301-09）资助项目
通讯作者: 郑璞     E-mail: zhengpu@jiangnan.edu.cn
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引用本文:

刘伟, 郑璞, 靳新娜. 阻断嗜乙酰乙酸棒杆菌乙酸合成途径对其在缺氧条件下产琥珀酸的影响[J]. 中国生物工程杂志, 2014, 34(9): 48-55.

LIU Wei, ZHENG Pu, JIN Xin-na. Effects of Disrupting Acetate Formation Pathways in Corynebacterium acetoacidophilum on Succinate Production Under Oxygen Deprivation. China Biotechnology, 2014, 34(9): 48-55.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20140908    或    https://manu60.magtech.com.cn/biotech/CN/Y2014/V34/I9/48


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