Quantitative analysis and engineering of fatty acid biosynthesis in E. coli
Quantitative analysis and engineering of fatty acid biosynthesis in E. coli
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DOI：
10.1016/j.ymben.2010.02.003
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发表时间：
2010-07-01
影响因子：
8.4
通讯作者：
Khosla, Chaitan
中科院分区：
文献类型：
--
作者：
Liu, Tiangang;Vora, Harmit;Khosla, Chaitan
脂肪酸是从可再生资源生物合成柴油的中心烃中间体。我们已经设计了一种大肠杆菌细胞系，其在补料分批发酵中产生4.5g/L/天的总脂肪酸。然而，在该细胞系中脂肪酸生物合成的进一步增强被证明是不可预测的。为了开发更可靠的工程策略，开发了一种无细胞系统，该系统能够直接定量研究E.杆菌使用该系统，脂肪酸合成对丙二酰辅酶A可用性的强依赖性和几个重要的现象，目的脂肪酸合成进行了验证。通过代谢工程菌株的产生和分析，证实了这种无细胞系统的结果。杆菌我们的定量研究结果突出了E.大肠杆菌脂肪酸生物合成途径，并针对蛋白质和代谢工程的具体步骤，以提高葡萄糖催化转化为生物柴油。(C)2010年爱思唯尔公司All rights reserved.
Fatty acids are central hydrocarbon intermediates in the biosynthesis of diesel from renewable sources. We have engineered an Escherichia coli cell line that produces 4.5g/L/day total fatty acid in a fed-batch fermentation. However, further enhancement of fatty acid biosynthesis in this cell line proved unpredictable. To develop a more reliable engineering strategy, a cell-free system was developed that enabled direct, quantitative investigation of fatty acid biosynthesis and its regulation in E. coli. Using this system, the strong dependence of fatty acid synthesis on malonyl-CoA availability and several important phenomena aim fatty acid synthesis were verified. Results from this cell-free system were confirmed via the generation and analysis of metabolically engineered strains of E. coli. Our quantitative findings highlight the enormous catalytic potential of the E. coli fatty acid biosynthetic pathway, and target specific steps for protein and metabolic engineering to enhance the catalytic conversion of glucose into biodiesel. (C) 2010 Elsevier Inc. All rights reserved.