Coupled ARTP and ALE strategy to improve anaerobic cell growth and succinic acid production by Escherichia coli
Coupled ARTP and ALE strategy to improve anaerobic cell growth and succinic acid production by Escherichia coli
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DOI：
10.1002/jctb.4633
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发表时间：
2016-03
影响因子：
3.4
通讯作者：
Jiangfeng Ma;Ming-ke Wu;Changqing Zhang;Ai-yong He;Xiang-Ping Kong;Ganlu Li;Ce Wei;M. Jiang
中科院分区：
文献类型：
--
作者：
Jiangfeng Ma;Ming-ke Wu;Changqing Zhang;Ai-yong He;Xiang-Ping Kong;Ganlu Li;Ce Wei;M. Jiang
大肠杆菌AFP 111，一种兼性厌氧菌株，在完全厌氧发酵期间在基本培养基中显示出非常低的细胞生长和琥珀酸生产速率。因此，采用大气压和室温等离子体（ARTP）和适应性实验室进化（ALE）策略来提高厌氧条件下的细胞生长和琥珀酸生产。结果经ARTP和ALE筛选获得一株突变株BER 208。在3L搅拌式生物反应器中培养时，BER 208的比生长速率是AFP 111的3.12倍。此外，它消耗40 g L-1葡萄糖，生产27.9 g L-1琥珀酸，葡萄糖利用率为0.54 g L-1 h-1，琥珀酸生产率为0.38 g L-1 h-1，分别是原始菌株的3.7倍和2.5倍。进一步的研究表明，该突变株的葡萄糖激酶和磷酸烯醇式丙酮酸羧激酶（PCK）活性分别比出发菌株提高了3.3倍和16倍。结论ARTP和ALE联合策略是获得性能提高的突变体的有效途径。© 2015化学工业协会
BACKGROUND Escherichia coli AFP111, a facultative anaerobic strain, shows a very low cell growth and succinic acid production rate in minimal medium during exclusively anaerobic fermentation. Therefore, atmospheric and room temperature plasma (ARTP) and adaptive laboratory evolution (ALE) strategies were adopted to improve cell growth and succinic acid production under anaerobic conditions. RESULTS A mutant, BER208, was obtained by ARTP and ALE used successively. When cultured in a 3 L stirred bioreactor, the specific growth rate of BER208 was increased 3.12-fold compared with AFP111. Also, it consumed 40 g L−1 glucose and produced 27.9 g L−1 succinic acid with a glucose utilization rate of 0.54 g L−1 h−1 and succinic acid productivity of 0.38 g L−1 h−1, which were 3.7-fold and 2.5-fold higher than those of the original strain. Further investigation showed that the activities of glucokinase and phosphoenolpyruvate carboxykinase (PCK) were increased 3.3-fold and 16-fold in the mutant compared with the original strain. CONCLUSION The combined strategy of ARTP and ALE was an effective approach to obtain mutants with improved performance. © 2015 Society of Chemical Industry