Enhancing succinic acid biosynthesis in Escherichia coli by engineering its global transcription factor, catabolite repressor/activator (Cra).
Enhancing succinic acid biosynthesis in Escherichia coli by engineering its global transcription factor, catabolite repressor/activator (Cra).
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通过改造大肠杆菌的全局转录因子、分解代谢物阻遏物/激活物 (Cra) 来增强大肠杆菌中的琥珀酸生物合成
DOI：
10.1038/srep36526
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发表时间：
2016-11-04
影响因子：
4.6
通讯作者：
Tang YJ
中科院分区：
文献类型：
--
作者：
Zhu LW;Xia ST;Wei LN;Li HM;Yuan ZP;Tang YJ
本研究旨在通过改造大肠杆菌全局转录因子CRA(分解代谢抑制因子/激活剂)来提高大肠杆菌琥珀酸的产量。随机突变文库通过容易出错的CRA聚合酶链式反应产生。经过重新筛选和突变位点整合，最好的突变菌株是唐1541，其最终琥珀酸浓度为79.8±3.1g/L，比空载体对照提高了22.8%。通过CRA突变，参与磷酸烯醇式丙酮酸(PEP)羧化和乙醛酸途径的基因和酶被直接或间接地激活。CRA与DNA相互作用的参数表明，CRA突变体与ACEBAK结合，从而激活了乙醛酸途径中涉及的基因，并进一步提高了琥珀酸合成，即使在其效应因子1，6-二磷酸果糖(FBP)存在的情况下也是如此。这表明，FBP对CRA的一些负面影响可能是通过提高CRA突变体与FBP的结合亲和力或改变CRA结构来抵消的。这项工作为理解琥珀酸生物合成的转录调控提供了有用的信息。
This study was initiated to improve E. coli succinate production by engineering the E. coli global transcription factor, Cra (catabolite repressor/activator). Random mutagenesis libraries were generated through error-prone PCR of cra. After re-screening and mutation site integration, the best mutant strain was Tang1541, which provided a final succinate concentration of 79.8 ± 3.1 g/L: i.e., 22.8% greater than that obtained using an empty vector control. The genes and enzymes involved in phosphoenolpyruvate (PEP) carboxylation and the glyoxylate pathway were activated, either directly or indirectly, through the mutation of Cra. The parameters for interaction of Cra and DNA indicated that the Cra mutant was bound to aceBAK, thereby activating the genes involved in glyoxylate pathway and further improving succinate production even in the presence of its effector fructose-1,6-bisphosphate (FBP). It suggested that some of the negative effect of FBP on Cra might have been counteracted through the enhanced binding affinity of the Cra mutant for FBP or the change of Cra structure. This work provides useful information about understanding the transcriptional regulation of succinate biosynthesis.
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影响因子：
3.6
作者：
RAMSEIER, TM;BLEDIG, S;SAIER, MH
通讯作者：
SAIER, MH
影响因子：
4.8
作者：
Chavarria, Max;Santiago, Cesar;de Lorenzo, Victor
通讯作者：
de Lorenzo, Victor
联合激活磷酸烯醇丙酮酸羧化酶和磷酸烯醇丙酮酸羧激酶以提高琥珀酸产量
影响因子：
4.4
作者：
Tan, Zaigao;Zhu, Xinna;Zhang, Xueli
通讯作者：
Zhang, Xueli
影响因子：
3.2
作者：
Bledig, SA;Ramseier, TM;Saier, MH
通讯作者：
Saier, MH
DOI：
10.1099/00221287-87-1-37
发表时间：
1975-01-01
期刊：
JOURNAL OF GENERAL MICROBIOLOGY
影响因子：
--
作者：
BENNETT, PM;HOLMS, WH
通讯作者：
HOLMS, WH