Resolving the TCA cycle and pentose-phosphate pathway of Clostridium acetobutylicum ATCC 824: Isotopomer analysis, in vitro activities and expression analysis.
Resolving the TCA cycle and pentose-phosphate pathway of Clostridium acetobutylicum ATCC 824: Isotopomer analysis, in vitro activities and expression analysis.
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解决乙梭状芽孢杆菌ATCC的TCA循环和五磷酸五磷酸途径824：同位素分析，体外活性和表达分析。
DOI：
10.1002/biot.201000282
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发表时间：
2011-03
影响因子：
4.7
通讯作者：
Antoniewicz MR
中科院分区：
文献类型：
--
作者：
Crown SB;Indurthi DC;Ahn WS;Choi J;Papoutsakis ET;Antoniewicz MR
溶剂型梭状芽孢杆菌是一类重要的能产生多种生物燃料的微生物。工程化梭状芽孢杆菌的瓶颈之一来自于中心代谢途径仍然知之甚少的事实。在这里，我们利用基于13C的同位素分析的力量来重新研究乙酰丁酸杆菌ATCC 824的中心代谢途径。我们使用[1，2-13C]葡萄糖，细胞内代谢产物的质谱分析和酶分析证明，乙酰丁酸杆菌有一个分裂的三氯乙酸循环，其中只有再柠檬酸合成酶通过柠檬酸参与α-酮戊二酸的产生。此外，我们发现α-酮戊二酸和富马酸之间没有碳交换，氧化戊糖-磷酸途径是不活跃的。对推测的Re-柠檬酸合成酶基因(CAC0970)及其操纵子以及所有糖酵解、戊糖-磷酸途径和TCA循环基因的动态基因表达分析确定了这些核心途径中的基因及其参与程度，这些核心途径支持这个工业有机体强大的初级代谢。
Solventogenic clostridia are an important class of microorganisms that can produce various biofuels. One of the bottlenecks in engineering clostridia stems from the fact that central metabolic pathways remain poorly understood. Here, we utilized the power of 13C-based isotopomer analysis to re-examine central metabolic pathways of C. acetobutylicum ATCC 824. We demonstrate using [1,2-13C]glucose, mass spectrometry analysis of intracellular metabolites, and enzymatic assays that C. acetobutylicum has a split TCA cycle where only Re-citrate synthase contributes to the production of α-ketoglutarate via citrate. Furthermore, we show that there is no carbon exchange between α-ketoglutarate and fumarate and that the oxidative pentose-phosphate pathway is inactive. Dynamic gene expression analysis of the putative Re-citrate synthase gene (CAC0970), its operon, and all glycolysis, pentose-phosphate pathway and TCA cycle genes identify genes and their degree of involvement in these core pathways that support the powerful primary metabolism of this industrial organism.
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影响因子：
8.4
作者：
Sillers, Ryan;Chow, Alison;Papoutsakis, Eleftherios T.
通讯作者：
Papoutsakis, Eleftherios T.
影响因子：
3.8
作者：
Young, Jarney D.;Walther, Jason L.;Stephanopoulos, Gregory
通讯作者：
Stephanopoulos, Gregory
影响因子：
8.4
作者：
Antoniewicz, Maciek R.;Kraynie, David F.;Stephanopoulos, Gregory
通讯作者：
Stephanopoulos, Gregory
影响因子：
4.7
作者：
Salimi, Fahimeh;Zhuang, Kai;Mahadevan, Radhakrishnan
通讯作者：
Mahadevan, Radhakrishnan
影响因子：
3.8
作者：
Senger, Ryan S.;Papoutsakis, Eleftherios T.
通讯作者：
Papoutsakis, Eleftherios T.