An indole‐deficient        <i xmlns='http://www.wiley.com/namespaces/wiley'>Escherichia coliEscherichia coli        strain improves screening of cytochromes P450 for biotechnological applications

Brixius‐Anderko Simone; Hannemann Frank; Ringle Michael; Khatri Yogan; Bernhardt Rita

摘要

Abstract > Escherichia coli has developed into an attractive organism for heterologous cytochrome P450 production, but, in some cases, was restricted as a host in view of a screening of orphan cytochromes P450 or mutant libraries in the context of molecular evolution due to the formation of the cytochrome P450 inhibitor indole by the enzyme tryptophanase (TnaA). To overcome this effect, we disrupted the tnaA gene locus of E.?coli C43(DE3) and evaluated the new strain for whole‐cell substrate conversions with three indole‐sensitive cytochromes P450, myxobacterial CYP264A1, and CYP109D1 as well as bovine steroidogenic CYP21A2. For purified CYP264A1 and CYP21A2, the half maximal inhibitory indole concentration was determined to be 140 and 500?μM, which is within the physiological concentration range occurring during cultivation of E.?coli in complex medium. Biotransformations with C43(DE3)_? tnaA achieved a 30% higher product formation in the case of CYP21A2 and an even fourfold increase with CYP264A1 compared with C43(DE3) cells. In whole‐cell conversion based on CYP109D1, which converts indole to indigo, we could successfully avoid this reaction. Results in microplate format indicate that our newly designed strain is a suitable host for a fast and efficient screening of indole‐influenced cytochromes P450 in complex medium. </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”> <title type =“main”>抽象</ title> > escherichia coli </ i>已经发展成为异源细胞色素P450生产的有吸引力的生物体，但在某些情况下，考虑到孤儿中的筛选是孤儿中的筛选或突变文库的宿主，因为在分子演化的背景下由于细胞色素P450抑制剂的形成而受到筛选吲哚色氨酸酶（TNAA）吲哚。为了克服这种效果，我们破坏了 E.?COLI </ i> C43（DE3）的 TNAA </ i>基因座，并评估了具有三个吲哚敏感的全细胞基材转换的新菌株细胞色素P450，骨膜细菌CYP264A1和CYP109D1以及牛类体化CYP21A2。对于纯化的CYP264A1和CYP21A2，将半最大抑制吲哚浓度确定为140和500μm，其在复合介质中培养期间在培养期间发生的生理浓度范围内。具有C43（DE3）_的生物转化形式_？ TNAA </ i>在CYP21A2的情况下达到30％的产物形成，与CYP264A1相比，与C 4 3（DE3）细胞相比，甚至四倍。在基于CYP109D1的全细胞转化中，将吲哚转化为靛蓝，我们可以成功地避免这种反应。微孔板格式的结果表明，我们的新设计的菌株是一种合适的宿主，用于快速有效地筛选在复合介质中的吲哚影响的细胞色素P450。 </ p> </ 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-15016/'>《Biotechnology and Applied Biochemistry》</a> <b style="margin: 0 2px;">|</b><span>2017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=Brixius‐Anderko Simone&option=202" target="_blank" rel="nofollow">Brixius‐Anderko Simone;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Hannemann Frank&option=202" target="_blank" rel="nofollow">Hannemann Frank;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ringle Michael&option=202" target="_blank" rel="nofollow">Ringle Michael;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Khatri Yogan&option=202" target="_blank" rel="nofollow">Khatri Yogan;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Bernhardt Rita&option=202" target="_blank" rel="nofollow">Bernhardt Rita;</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>Department of BiochemistrySaarland UniversitySaarbrücken Germany;</p> <p>Department of BiochemistrySaarland UniversitySaarbrücken Germany;</p> <p>Department of BiochemistrySaarland UniversitySaarbrücken Germany;</p> <p>Department of BiochemistrySaarland UniversitySaarbrücken Germany;</p> <p>Department of BiochemistrySaarland UniversitySaarbrücken Germany;</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><a href="https://www.zhangqiaokeyan.com/clc/177.html" title="生物化学">生物化学;</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=cytochrome P450&option=203" rel="nofollow">cytochrome P450;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=E.?coli&option=203" rel="nofollow">E.?coli;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=gene disruption&option=203" rel="nofollow">gene disruption;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=indole&option=203" rel="nofollow">indole;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=screening&option=203" rel="nofollow">screening;</a> </p> <div class="translation"> 机译：细胞色素p450;E.？Coli;基因破坏;吲哚;筛选; </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 class="active" >外文文献</li> <li >中文文献</li> <li >专利</li> </ul> </div> <div class="similarity_details"> <ul > <li> <div> <b>1. </b><a class="enjiyixqcontent" 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Andrew&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Hausrath Andrew,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Sagermann Martin&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Sagermann Martin,</a> <span>1997</span> </span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：通过使用细胞色素缺乏硼的大肠杆菌改进的ECF1和ECF1F0纯化有助于这些复合物的结晶</span> </p> </li> <li> <div> <b>11. </b><a class="enjiyixqcontent" href="/ntis-science-report_ad_thesis/020711484457.html">Synthesis of Aerobactin and a 76,000-Dalton Iron-Regulated Outer Membrane Protein by Escherichia coli K-12-Shigella flexneri Hybrids and by Enteroinvasive Strains of Escherichia coli</a> <b>[R] </b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Griffiths, E.&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Griffiths, E.,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Stevenson, P.&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Stevenson, P.,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Hale, T. L.&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Hale, T. L.,</a> <span>1985</span> </span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：大肠埃希氏菌K-12-shigella flexneri杂交种和大肠杆菌肠道侵袭菌合成aerobactin和76,000-Dalton铁调节的外膜蛋白</span> </p> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/academic-journal-cn_zoological-research_thesis/0201290688706.html">Clonal spread of Escherichia coli O101:H9-ST10 and O101:H9-ST167 strains carrying fosA3 and blaCTX-M-14 among diarrheal calves in a Chinese farm,with Australian Chroicocephalus as the possible origin of E.coli O101:H9-ST10</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Wan-Yun He&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . Wan-Yun He</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Jian-Hua Liu&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Jian-Hua Liu</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Xing-Xing Zhang&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Xing-Xing Zhang</a> <span> <a href="/journal-cn-5045/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 动物学研究 </a> </span> <span> . 2021</span><span>,第004期</span> </span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/academic-journal-cn_world-journal-gastroenterology-english_thesis/0201296588608.html">Intestinal-borne dermatoses significantly improved by oral application of Escherichia coli Nissle 1917</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Elina Manzhalii&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . Elina Manzhalii</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Daniel Hornuss&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Daniel Hornuss</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Wolfgang Stremmel&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Wolfgang Stremmel</a> <span> <a href="/journal-cn-52914/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 世界胃肠病学杂志：英文版 </a> </span> <span> . 2016</span><span>,第23期</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/06130400497340.html">GENE-THERAPEUTIC DNA-VECTOR BASED ON GENE-THERAPEUTIC DNA-VECTOR VTVAF17, CARRYING TARGET GENE SELECTED FROM GROUP OF GENES ANG, ANGPT1, VEGFA, FGF1, HIF1Α, HGF, SDF1, KLK4, PDGFC, PROK1, PROK2 TO INCREASE EXPRESSION LEVEL OF SAID TARGET GENES, METHOD FOR PRODUCTION AND USE THEREOF, STRAIN ESCHERICHIA COLI SCS110-AF/VTVAF17-ANG, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-ANGPT1, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-VEGFA, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-FGF1, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-HIF1Α, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-HGF, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-SDF1, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-KLK4, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-PDGFC, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-PROK1, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-PROK2, CARRYING GENE-THERAPEUTIC DNA VECTOR, METHOD FOR PRODUCTION THEREOF, METHOD FOR INDUSTRIAL PRODUCTION OF GENE-THERAPEUTIC DNA VECTOR</a> <b>[P]</b> . <span> 外国专利：  RU2730664C2 </span> <span> . 2020-08-24</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：基于基因治疗DNA载体VTVAF17的基因治疗DNA载体，携带选自基因ANG，ANGPT1，VEGFA，FGF1，HIF1α，HGF，SDF1，KLK4，PDGFC，PROK1，PROK2表达的靶基因所述的靶标基因，其生产和使用方法，应变大肠埃希氏菌SCS110-AF / VTVAF17-ANG或大肠埃希氏菌SCS110-AF / VTVAF17-ANGPT1或大肠埃希氏菌SCS110-AF / VTVAF17-VEGFA或大肠埃希氏菌SCS110-AF / VTVAF17-FGF1，或大肠埃希氏菌SCS110-AF / VTVAF17-HIF1A，或大肠埃希氏菌COLI SCS110-AF / VTVAF17-HGF，或大肠埃希氏菌SCS110-AF / VTVAF17-SDF1，或大肠埃希氏菌SCS110-AF / VTVAF17-KLK4，大肠埃希氏菌SCS110-AF / VTVAF17-PDGFC或大肠埃希氏菌SCS110-AF / VTVAF17-PROK2，携带基因-治疗性DNA载体，生产方法，工业生产方法-治疗性DNA载体 </span> </p> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/06130400493335.html">GENE-THERAPEUTIC DNA VECTOR BASED ON THE GENE-THERAPEUTIC DNA VECTOR GDTT1_8NAS12, CARRYING THE TARGET GENE SELECTED FROM A GROUP OF GENES DDC, IL10, IL13, IFNB1, TNFRSF4, TNFSF10, BCL2, HGF, IL2 TO INCREASE THE EXPRESSION LEVEL OF SAID TARGET GENES, A METHOD FOR PRODUCTION AND USE THEREOF, A STRAIN ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-DDC OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-IL10 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-IL13 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-IFNB1 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-TNFRSF4 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-TNFSF10 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-BCL2 OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-HGF OR ESCHERICHIA COLI JM110-NAS/GDTT1_8NAS12-IL2, CARRYING A GENE-THERAPEUTIC DNA VECTOR, METHOD FOR PRODUCTION THEREOF, A METHOD FOR INDUSTRIAL PRODUCTION OF A GENE-THERAPEUTIC DNA VECTOR</a> <b>[P]</b> . <span> 外国专利：  RU2734726C1 </span> <span> . 2020-10-22</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：基于基因治疗DNA矢量GDTT1_8NAS12的基因治疗DNA矢量，携带从一组基因中选择的目标基因DDC，IL10，IL13，IFNB1，TNFRSF4，TNFSF10，BCL2，HGF，IL2可以增加表达水平基因，一种生产和使用其的方法，应变大肠埃希氏菌JM110-NAS / GDTT1_8NAS12-DDC或大肠埃希氏菌JM110-NAS / GDTT1_8NAS12-IL10或大肠埃希氏菌JM110-NAS / GDTT1_8NAS12-IL13或大肠埃希氏菌COLI JM110-NAS / GDTT1_8NAS12-IL13 IFNB1或ESCHERICHIA COLI JM110-NAS / GDTT1_8NAS12-TNFRSF4或ESCHERICHIA COLI JM110-NAS / GDTT1_8NAS12-TNFSF10或ESCHERICHIA COLI JM110-NAS / GDTT1_8NAS12-BCL2或ESCHERICHIA COLI JM110-NAS / GDTT1_8NAS12 IL2，携带基因治疗性DNA载体，其生产方法，工业生产基因治疗性DNA载体的方法 </span> </p> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130400537005.html">Gene therapeutic DNA vector based on VTvaf17 gene therapeutic DNA vector carrying a target gene selected from the group of genes ANG, ANGPT1, VEGFA, FGF1, HIF1α, HGF, SDF1, KLK4, PDGFC, PROK1, PROK2 to increase the expression level of these target genes, method its preparation and use, Escherichia coli strain SCS110-AF / VTvaf17-ANG or Escherichia coli SCS110-AF / VTvaf17-ANGPT1 or Escherichia coli SCS110-AF / VTvaf17-VEGFA or Escherichia coli SCS110-AF / VTvaf17-Fichia-SC110 AF / VTvaf17-HIF1α or Escherichia coli SCS110-AF / VTvaf17-HGF or Escherichia coli SCS110-AF / VTvaf17-SDF1 or Escherichia coli SCS110-AF / VTvaf17-KLK4 or Escherichia coli SCS110-AF / VTviFi10 SCF110 AF / VTvaf17-PROK1 or Escherichia coli SCS110-AF / VTvaf17-PROK2 carrying a gene therapy DNA vector, a method for its preparation, a method for the industrial production of a gene therapy DNA vector</a> <b>[P]</b> . <span> 外国专利：  RU2018147085A </span> <span> . 2020-06-29</span> </div> <p class="zwjiyix translation" style="max-width: initial;height: auto;word-break: break-all;white-space: initial;text-overflow: initial;overflow: initial;"> <span>机译：基于VTvaf17基因治疗DNA载体的基因治疗DNA载体，其携带选自ANG，ANGPT1，VEGFA，FGF1，HIF1α，HGF，SDF1，KLK4，PDGFC，PROK1，PROK2的靶基因以增加这些靶标的表达水平基因，方法的制备和使用，大肠杆菌菌株SCS110-AF / VTvaf17-ANG或大肠杆菌SCS110-AF / VTvaf17-ANGPT1或大肠杆菌SCS110-AF / VTvaf17-VEGFA或大肠杆菌SCS110-AF / VTvaf17-Fichia-SC110 AF /VTvaf17-HIF1α或大肠杆菌SCS110-AF / VTvaf17-HGF或大肠杆菌SCS110-AF / VTvaf17-SDF1或大肠杆菌SCS110-AF / VTvaf17-KLK4或大肠杆菌SCS110-AF / VTviFi10 S1携带基因治疗DNA载体的大肠杆菌SCS110-AF / VTvaf17-PROK2，其制备方法，工业生产基因治疗DNA载体的方法 </span> </p> </li> </ul> </div> </div> </div> <div class="theme cardcommon" style="overflow: auto;display:none"> <h3 class="all_title" id="enpatent55">相关主题</h3> <ul id="subject"> </ul> </div> </div> </div> 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An indole‐deficient Escherichia coliEscherichia coli strain improves screening of cytochromes P450 for biotechnological applications

摘要