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Application of fermentation process control to increase l‐tryptophan production in Escherichia coli

机译：发酵过程控制在大肠杆菌中提高L-色氨酸生产的应用

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Abstract > In this study, process engineering and process control were applied to increase the production of <sc>l</sc> ‐tryptophan using <fc> <fi>Escherichia coli</fi> </fc> D <fi>mtr</fi> / <fi>pta</fi> ‐Y. Different dissolved oxygen (DO) and pH control strategies were applied in <sc>l</sc> ‐tryptophan production. DO and pH were maintained at [20% (0–20?hr); 30% (20–40?hr)] and [7.0 (0–20?hr), 6.5 (20–40?hr)], respectively, which increased <sc>l</sc> ‐tryptophan production, glucose conversion percentage [g ( <sc>l</sc> ‐tryptophan)/g (glucose)], and transcription levels of key genes for tryptophan biosynthesis and tryptophan biosynthesis flux, and decreased the accumulation of acetate and transcription levels of genes related to acetate synthesis and acetate synthesis flux. Using <fc> <fi>E. coli</fi> </fc> D <fi>mtr</fi> / <fi>pta</fi> ‐Y with optimized DO [20% (0–20?hr); 30% (20–40?hr)] and pH [7.0 (0–20?hr), 6.5 (20–40?hr)] values, the highest <sc>l</sc> ‐tryptophan production (52.57?g/L) and glucose conversion percentage (20.15%) were obtained. The <sc>l</sc> ‐tryptophan production was increased by 26.58%, the glucose conversion percentage was increased by 22.64%, and the flux of tryptophan biosynthesis was increased to 21.5% compared with different conditions for DO [50% (0–20?hr), 20% (20–40?hr)] and pH [7.0]. </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 Type =“Main”XML：Lang =“en”> <标题类型=“main”>抽象</ title> > 在这项研究中，应用工艺工程和过程控制来增加生产 <sc> l </ sc> -Treptophan使用 <FC> <fi>大肠杆菌</ fi> </ FC> D. <fi> mtr </ fi> / <fi> pta </ fi> - 那。应用不同的溶解氧（DO）和pH控制策略 <sc> l </ sc> -tryptophan生产。 DO和pH维持在[20％（0-20-小时）; 30％（20-40？HR）]和[7.0（0-20？HR），6.5（20-40？HR）]增加 <sc> l </ sc> - 生成，葡萄糖转化百分比[g（ <sc> l </ sc> -Tryptophan）/ g（葡萄糖）]和色氨酸生物合成和色氨酸生物合成通量的关键基因的转录水平，并降低了醋酸酯合成和醋酸酯合成通量相关的基因的积累和转录水平。使用 <FC> <fi> e。 COLI </ FI> </ FC> D. <fi> mtr </ fi> / <fi> pta </ fi> - 很优化DO [20％（0-20？HR）; 30％（20-40？HR）]和pH [7.0（0-20〜HR），6.5（20-40？HR）]值，最高 <sc> l </ sc> - 获得 - 基团生产（52.57〜11 / L）和葡萄糖转化率（20.15％）。这 <sc> l </ sc> - 活跃产量增加26.58％，葡萄糖转化率提高了22.64％，色氨酸生物合成的通量增加到21.5％，与DO的不同条件[50％（0-20〜20℃），20％（ 20-40？hr）和pH [7.0]。 </ p> </摘要> </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-15005/'>《Biotechnology Progress》</a> <b style="margin: 0 2px;">|</b><span>2020年第2期</span><b style="margin: 0 2px;">|</b><span>共10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=Zhao Chunguang&option=202" target="_blank" rel="nofollow">Zhao Chunguang;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Fang Haitian&option=202" target="_blank" rel="nofollow">Fang Haitian;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Wang Jing&option=202" target="_blank" rel="nofollow">Wang Jing;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Zhang Shasha&option=202" target="_blank" rel="nofollow">Zhang Shasha;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Zhao Xiubao&option=202" target="_blank" rel="nofollow">Zhao Xiubao;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Li Zengliang&option=202" target="_blank" rel="nofollow">Li Zengliang;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Lin Chuwen&option=202" target="_blank" rel="nofollow">Lin Chuwen;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Shen Zhiqiang&option=202" target="_blank" rel="nofollow">Shen Zhiqiang;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Cheng Likun&option=202" target="_blank" rel="nofollow">Cheng Likun;</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>Key Laboratory of Fermentation EngineeringShandong Binzhou Animal Science and Veterinary Medicine AcademyBinzhou China;</p> <p>School of AgricultureNingxia University Ningxia Eppen Biotech Co. LtdYinchuan China;</p> <p>Key Laboratory of Fermentation EngineeringShandong Binzhou Animal Science and Veterinary Medicine AcademyBinzhou China;</p> <p>Key Laboratory of Fermentation EngineeringShandong Binzhou Animal Science and Veterinary Medicine AcademyBinzhou China;</p> <p>Key Laboratory of Fermentation EngineeringShandong Binzhou Animal Science and Veterinary Medicine AcademyBinzhou China;</p> <p>Shandong Research Center of High Cell Density Fermentation and Efficient Expression TechnologyShandong Lvdu Bio‐science and Technology Co. LtdBinzhou China;</p> <p>Key Laboratory of Fermentation EngineeringShandong Binzhou Animal Science and Veterinary Medicine AcademyBinzhou China;</p> <p>Key Laboratory of Fermentation EngineeringShandong Binzhou Animal Science and Veterinary Medicine AcademyBinzhou China;</p> <p>Key Laboratory of Fermentation EngineeringShandong Binzhou Animal Science and Veterinary Medicine AcademyBinzhou 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/15.html" title="生物科学">生物科学;</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=Escherichia coli&option=203" rel="nofollow">Escherichia coli;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=l‐tryptophan&option=203" rel="nofollow">l‐tryptophan;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=metabolic flux&option=203" rel="nofollow">metabolic flux;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=process control&option=203" rel="nofollow">process control;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=transcription&option=203" rel="nofollow">transcription;</a> </p> <div class="translation"> 机译：大肠杆菌;L-色氨酸;代谢助焊剂;过程控制;转录; </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"> 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<span>机译：大肠杆菌中乙酸盐生物合成途径的基因修饰和细胞回收技术的实施以增加L-色氨酸的产生</span> </p> </li> <li> <div> <b>11. </b><a class="enjiyixqcontent" href="/open-access_resources_thesis/01000105516285.html ">Gene modification of the acetate biosynthesis pathway in Escherichia coli and implementation of the cell recycling technology to increase L-tryptophan production.</a> <b>[O] </b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Qingyang Xu&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Qingyang Xu,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Fang Bai&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Fang Bai,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ning Chen&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Ning Chen,</a> <span>2017</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：在大肠杆菌中基因修饰乙酸酯生物合成途径并实施细胞再循环技术以增加L-色氨酸的产生。</span> </p> </li> <li> <div> <b>12. </b><a class="enjiyixqcontent" href="/ntis-science-report_ad_thesis/02071653174.html">1000-L Scale-Up Fermentation of Escherichia Coli Containing PVSEOP7 for Production of Organophosphorus Hydrolase</a> <b>[R] </b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Lukens, D. C. &option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Lukens, D. C. ,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Rastogi, V. K. &option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Rastogi, V. K. ,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Cheng, T. &option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Cheng, T. ,</a> <span>2003</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：含有pVsEOp7的大肠杆菌1000-L放大发酵生产有机磷水解酶</span> </p> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/academic-journal-cn_carcinogenesis-teratogenesis-mutagenesis_thesis/0201285799305.html">The Mutagenesis In Tryptophan-requiring Auxotrophs of Escherichia Coli Induced By Tritium</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . </a> <span> <a href="/journal-cn-6199/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 癌变．畸变．突变 </a> </span> <span> . 1991</span><span>,第0S1期</span> </span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/academic-conference-cn_meeting-9099_thesis/020222283989.html">Study on volatile fatty acids production from rice stalk during the dry fermentation process and development of microbial populations</a> <b>[C]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=ZHAO Guang&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . ZHAO Guang</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=WEI Li&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,WEI Li</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=MA Fang&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,MA Fang</a> <span> <a href="/conference-cn-9099/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 第五届全国博士生会议暨环境科学与工程新理论、新技术学术研讨会 </a> <span> <span> . 2011</span> </span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/academic-degree-domestic_mphd_thesis/020312904087.html">纳米银对Escherichia coli细胞活性及可培养性的胁迫机制研究</a> <b>[A] </b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=卢雪蓉&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 卢雪蓉</a> <span> . 2018</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/061205128252.html">一种大肠杆菌Escherichia coli ETEC BE-311菌株及其应用</a> <b>[P]</b> . <span> 中国专利： CN112625989B </span> <span> . 2022.05.17</span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/061205128611.html">大肠杆菌Escherichia coli ETEC BE-311-faeG及应用</a> <b>[P]</b> . <span> 中国专利： CN112522172B </span> <span> . 2022.05.17</span> </div> </li> <li> <div> <b>3. </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>4. </b><a class="enjiyixqcontent" href="/patent-detail/06130400507597.html">GENE-THERAPY DNA-VECTOR BASED ON GENE-THERAPY DNA-VECTOR VTVAF17, CARRYING TARGET GENE SELECTED FROM GROUP OF GENES BMP-2, BMP-7, LMP-1, NELL-1 TO INCREASE EXPRESSION LEVEL OF THESE TARGET GENES, METHOD FOR PRODUCTION AND APPLICATION THEREOF, STRAIN ESCHERICHIA COLI SCS110-AF/VTVAF17-BMP-2, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-BMP-7, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-LMP-1, OR ESCHERICHIA COLI SCS110-AF/VTVAF17-NELL-1, CARRYING GENE-THERAPY DNA VECTOR, METHOD FOR PRODUCTION THEREOF, METHOD FOR INDUSTRIAL PRODUCTION OF GENE-THERAPY DNA VECTOR</a> <b>[P]</b> . <span> 外国专利：  RU2720521C1 </span> <span> . 2020-04-30</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，携带选自基因组BMP-2，BMP-7，LMP-1，NELL-1的靶基因以增加这些靶基因的表达水平，方法应变大肠埃希氏菌SCS110-AF / VTVAF17-BMP-2或大肠埃希氏菌SCS110-AF / VTVAF17-BMP-7或大肠埃希氏菌SCS110-AF / VTVAF17-LMP-1或大肠埃希氏菌SCS110-AF的生产和应用/ VTVAF17-NELL-1，携带基因治疗DNA矢量，其制备方法，工业生产基因治疗DNA矢量的方法 </span> </p> </li> <li> <div> <b>5. </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> </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> </div> <div class="right rightcon"> <div class="details_img cardcommon clearfix" style="margin-bottom: 10px;display:none;" > </div> </div> </div> <div id="thesis_get_original1" class="downloadBth" style="bottom: 19px;z-index: 999;" onclick="ywcd('0704028287933','4',7,2,1,'',this,24)" class="delivery" prompt="010401" title="通过人工服务将文献原文发送至邮箱" >获取原文</div> <div class="journalsub-pop-up" style="display: none"> <div class="journal-sub"> <h2>期刊订阅</h2> <img src="https://cdn.zhangqiaokeyan.com/img/loginclose.png" alt="" onclick="$('.journalsub-pop-up').hide()"> <p class="pardon">抱歉，该期刊暂不可订阅，敬请期待！</p> <p class="current">目前支持订阅全部北京大学中文核心（2020）期刊目录。</p> <div style="display: flex;margin-top: 69px;justify-content: 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