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首页> 外文期刊>Biotechnology and Bioengineering >Engineering Escherichia coliEscherichia coli for malate production by integrating modular pathway characterization with CRISPRi‐guided multiplexed metabolic tuning
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Engineering Escherichia coliEscherichia coli for malate production by integrating modular pathway characterization with CRISPRi‐guided multiplexed metabolic tuning

机译：工程 eScherichia coli 通过将模块化路径表征与CRISPRI-Guided Medopled Metabolic调整集成模块化路径表征来实现母体生产的大肠杆菌

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Abstract <section xml:id="bit26486-sec-0001" numbered="no"> > The application of rational design in reallocating metabolic flux to overproduce desired chemicals is always restricted by the native regulatory network. Here, we demonstrated that in vitro modular pathway optimization combined with in vivo multiplexed combinatorial engineering enables effective characterization of the bottleneck of a complex biosynthetic cascade and improves the output of the engineered pathway. As a proof of concept, we systematically identified the rate‐limiting step of a five‐gene malate biosynthetic pathway by combinatorially tuning the enzyme loads of a reconstituted biocatalytic reaction in a cell‐free system. Using multiplexed CRISPR interference, we subsequently eliminated the metabolic constraints by rationally assigning an optimal gene expression pattern for each pathway module. The present engineered strain Escherichia coli B0013‐47 exhibited a 2.3‐fold increase in malate titer compared with that of the parental strain, with a yield of 0.85?mol/mol glucose in shake‐flask culture and titer of 269?mM (36?g/L) in fed‐batch cultivation. The strategy reported herein represents a powerful method for improving the efficiency of multi‐gene pathways and advancing the success of metabolic engineering. </section> </abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> <div class="translation abstracttxt"> <span class="zhankaihshouqi fivelineshidden" id="abstract"> <span>机译：</span><Abstract XMLNS =“http://www.wiley.com/namespaces/wiley”type =“main”xml：lang =“en”> <title type =“main”>抽象</ title> <section XML：ID =“Bit26486-SEC-0001”编号=“否”> >在重新分配代谢通量的原理设计中的应用始终受到原生监管网络的预付费化学品。这里，我们证明，在体外模块化途径优化与体内多路复用组合工程结合的优化使得能够有效地表征复杂的生物合成级联的瓶颈并改善了工程路径的输出。作为概念证明，我们通过组合在无细胞系统中组合重构的生物催化反应的酶负荷来系统地确定了五基丙酸盐生物合成途径的速率限制步骤。使用多路复用的CRISPR干扰，我们随后通过合理分配每个途径模块的最佳基因表达模式来消除代谢约束。本工程化菌株大肠杆菌Coli </ i> B0013-47与亲本菌株相比，苹果酸盐滴度的增加2.3倍，产率为0.85Ω摩尔/摩尔葡萄糖中的羊瓶培养和滴度在FED分批栽培中269毫米（36？g / l）。本文报道的策略是提高多基因途径效率并推进代谢工程成功的强大方法。 </ p> </ section> </ abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> </div> <div class="record"> <h2 class="all_title" id="enpatent33" >著录项</h2> <ul> <li> <span class="lefttit">来源</span> <div style="width: 86%;vertical-align: text-top;display: inline-block;"> <a href='/journal-foreign-15017/'>《Biotechnology and Bioengineering》</a> <b style="margin: 0 2px;">|</b><span>2018年第3期</span><b style="margin: 0 2px;">|</b><span>共12页</span> </div> </li> <li> <div class="author"> <span class="lefttit">作者</span> <p id="fAuthorthree" class="threelineshidden zhankaihshouqi"> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Gao Cong&option=202" target="_blank" rel="nofollow">Gao Cong;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Wang Shihui&option=202" target="_blank" rel="nofollow">Wang Shihui;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Hu Guipeng&option=202" target="_blank" rel="nofollow">Hu Guipeng;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Guo Liang&option=202" target="_blank" rel="nofollow">Guo Liang;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Chen Xiulai&option=202" target="_blank" rel="nofollow">Chen Xiulai;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Xu Peng&option=202" target="_blank" rel="nofollow">Xu Peng;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Liu Liming&option=202" target="_blank" rel="nofollow">Liu Liming;</a> </p> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zkzz" style="display: none;">展开▼</span> </div> </li> <li> <div style="display: flex;"> <span class="lefttit">作者单位</span> <div style="position: relative;margin-left: 3px;max-width: 639px;"> <div class="threelineshidden zhankaihshouqi" id="fOrgthree"> <p>State Key Laboratory of Food Science and TechnologyJiangnan UniversityWuxi China;</p> <p>State Key Laboratory of Food Science and TechnologyJiangnan UniversityWuxi China;</p> <p>State Key Laboratory of Food Science and TechnologyJiangnan UniversityWuxi China;</p> <p>State Key Laboratory of Food Science and TechnologyJiangnan UniversityWuxi China;</p> <p>State Key Laboratory of Food Science and TechnologyJiangnan UniversityWuxi China;</p> <p>Chemical Biochemical and Environmental EngineeringUniversity of Maryland Baltimore CountyBaltimore Maryland;</p> <p>State Key Laboratory of Food Science and TechnologyJiangnan UniversityWuxi China;</p> </div> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zhdw" style="display: none;">展开▼</span> </div> </div> </li> <li > <span class="lefttit">收录信息</span> <span style="width: 86%;vertical-align: text-top;display: inline-block;"></span> </li> <li> <span class="lefttit">原文格式</span> <span>PDF</span> </li> <li> <span class="lefttit">正文语种</span> <span>eng</span> </li> <li> <span class="lefttit">中图分类</span> <span><a href="https://www.zhangqiaokeyan.com/clc/180.html" title="生物工程学（生物技术）">生物工程学（生物技术）;</a></span> </li> <li class="antistop"> <span class="lefttit">关键词</span> <p style="width: 86%;vertical-align: text-top;"> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=CRISPRi&option=203" rel="nofollow">CRISPRi;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=glyoxylate cycle&option=203" rel="nofollow">glyoxylate cycle;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=in vitro modular optimization&option=203" rel="nofollow">in vitro modular optimization;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=multiplexed combinatorial regulation&option=203" rel="nofollow">multiplexed combinatorial regulation;</a> </p> <div class="translation"> 机译：CRISPRI;乙醛循环;体外模块化优化;多路复用组合调节; </div> </li> </ul> </div> </div> <div class="literature cardcommon"> <div class="similarity "> <h3 class="all_title" id="enpatent66">相似文献</h3> <div class="similaritytab clearfix"> <ul> <li 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