Improved Production of Triostin A in Engineered Escherichia coli with Furnished Quinoxaline Chromophore by Design of Experiments in Small‐Scale Culture
Improved Production of Triostin A in Engineered Escherichia coli with Furnished Quinoxaline Chromophore by Design of Experiments in Small‐Scale Culture
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DOI：
10.1021/bp070298y
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发表时间：
2008
影响因子：
2.9
通讯作者：
Alex P Praseuth;Mike B. Praseuth;H. Oguri;H. Oikawa;Kenji Watanabe;Clay C. C. Wang-Clay-C.-C.-Wang-152744828
中科院分区：
文献类型：
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作者：
Alex P Praseuth;Mike B. Praseuth;H. Oguri;H. Oikawa;Kenji Watanabe;Clay C. C. Wang-Clay-C.-C.-Wang-152744828
使用工程化大肠杆菌 (E. coli) 开发了抗肿瘤药物 triostin A 的高效生产。该细菌拥有 15 个编码完整生物合成蛋白的基因，这些基因是从拉萨链霉菌中鉴定和克隆的。在这项研究中，随着外源喹喔啉-2-羧酸（QXC）的引入，三唑菌素 A 的产量显着增加了 20 倍以上，即 13 mg/L。QXC 是三唑菌素 A 生物合成的推测起始单位。相反，通过排除外源 QXC 的高细胞密度补料分批发酵从头生产三唑菌素 A，其中含有适量的抗肿瘤剂。通过小规模培养实现了生物活性分子的显着生产，并通过液相色谱-质谱仪完成了产品的定量分析。这个简单而快速的系统可以轻松地为我们提供有价值的信息，以最大限度地提高生产滴度。我们的完全异源生产系统也为未来使用大肠杆菌通过可耐受的前体定向生物合成生成新型生物活性化合物奠定了基础。
Proficient production of the antitumor agent triostin A was developed using engineered Escherichia coli ( E. coli). The bacterium played host to 15 genes that encode integral biosynthetic proteins which were identified and cloned from Streptomyces lasaliensis. In this study, triostin A production was dramatically increased by more than 20‐fold, 13 mg/L, with the introduction of exogenous quinoxaline‐2‐carboxylic acid (QXC), the speculative starting unit for biosynthesis of triostin A. Conversely, de novo production of triostin A by means of high cell density fed‐batch fermentation that is exclusive of exogenous QXC bore a modest amount of the antitumor agent. Noteworthy production of the biologically active molecule was achieved with small‐scale cultivation and quantitative analysis of the product was accomplished with a liquid chromatography‐mass spectrometer. This simple and speedy system could easily provide us with valuable information for maximizing the production titer. Our entirely heterologous production system also establishes a basis for the future use of E. coli for generation of novel bioactive compounds through tolerable precursor‐directed biosynthesis.