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Transcriptional engineering of the glyceraldehyde‐3‐phosphate dehydrogenase promoter for improved heterologous protein production in Pichia pastorisPichia pastoris

机译：甘油醛-3-磷酸脱氢酶启动子改善 pichia pastoris Pichia Pastoris的转录工程

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ABSTRACT <section xml:id="bit26363-sec-0001"> > The constitutive glyceraldehyde‐3‐phosphate dehydrogenase promoter (P <sub> GAP </sub> ), which is one of the benchmark promoters of Pichia pastoris , was analyzed in terms of putative transcription factor binding sites. We constructed a synthetic library with distinct regulatory properties through deletion and duplication of these putative transcription factor binding sites and selected transcription factor (TF) genes were overexpressed or deleted to understand their roles on heterologous protein production. Using enhanced green fluorescent protein, an expression strength in a range between 0.35‐ and 3.10‐fold of the wild‐type P <sub> GAP </sub> was obtained. Another model protein, recombinant human growth hormone was produced under control of selected promoter variants and 1.6‐ to 2.4‐fold higher product titers were reached compared to wild‐type P <sub> GAP </sub> . In addition, a GAL4‐like TF was found to be a crucial factor for the regulation of P <sub> GAP </sub> , and its overexpression enhanced the heterologous protein production considerably (up to 2.2‐fold compared to the parental strain). The synthetic P <sub> GAP </sub> library generated enabled us to investigate the different putative transcription factors which are responsible for the regulation of P <sub> GAP </sub> under different growth conditions, ergo recombinant protein production under P <sub> GAP </sub> . Biotechnol. Bioeng. 2017;114: 2319–2327. ? 2017 Wiley Periodicals, Inc. </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 =“bit26363-sec-0001”> >本构甘油醛-3-磷酸脱氢酶启动子（p <sub> 间隙</ i> </ sub>），这是<i的基准启动子之一>在推定的转录因子结合位点分析Pichia Pastoris </ i>。我们通过缺失和重复这些推定的转录因子结合位点和选择的转录因子（TF）基因过表达或删除，以了解其对异源蛋白质产生的作用，以制造具有不同调节性能的合成文库。使用增强的绿色荧光蛋白，得到0.35-和3.10倍的野生型P <sub>间隙</ i> </ sub>之间的表达强度。另一种模型蛋白质，重组人体生长激素是在选择的促进剂变体的控制下产生，与野生型P <Sub> <I>间隙相比，达到1.6-2.4倍的产品滴度</ i> </ sub>。另外，发现 Gal 4样ΔTF是对P <sub>间隙</ i> </ sub>调节的关键因素，其过表达增强了异源蛋白质生产大幅度（与家长菌株相比高达2.2倍）。合成的P <sub> 间隙</ i> </ sub>库使我们能够研究对P <sub>间隙的调节负责的不同推定转录因子</ i> </亚>在不同的生长条件下，在P <sub>间隙</ i> </ sub>下的ERGO重组蛋白质产生。 Biotechnol。生物。 2017; 114：2319-2327。还2017 Wiley期刊，Inc。</ 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>2017年第10期</span><b style="margin: 0 2px;">|</b><span>共9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=Ata ?zge&option=202" target="_blank" rel="nofollow">Ata ?zge;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Prielhofer Roland&option=202" target="_blank" rel="nofollow">Prielhofer Roland;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Gasser Brigitte&option=202" target="_blank" rel="nofollow">Gasser Brigitte;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Mattanovich Diethard&option=202" target="_blank" rel="nofollow">Mattanovich Diethard;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=?al?k P?nar&option=202" target="_blank" rel="nofollow">?al?k P?nar;</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 BiotechnologyMiddle East Technical UniversityAnkara 06800 Turkey;</p> <p>Department of BiotechnologyUniversity of Natural Resources and Life SciencesVienna Austria;</p> <p>Department of BiotechnologyUniversity of Natural Resources and Life SciencesVienna Austria;</p> <p>Department of BiotechnologyUniversity of Natural Resources and Life SciencesVienna Austria;</p> <p>Department of BiotechnologyMiddle East Technical UniversityAnkara 06800 Turkey;</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=Pichia pastoris&option=203" rel="nofollow">Pichia pastoris;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=GAP&option=203" rel="nofollow">GAP;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=promoter library&option=203" rel="nofollow">promoter library;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=transcriptional engineering&option=203" rel="nofollow">transcriptional engineering;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=transcription factor&option=203" rel="nofollow">transcription factor;</a> </p> <div class="translation"> 机译：Pichia Pastoris;Gap;启动子文库;转录工程;转录因子; </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" href="/journal-foreign-detail/0704021881866.html">Transcriptional 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2008</span><span>,第4期</span> </span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/academic-journal-cn_food-research-development_thesis/020123435648.html">基因工程菌Pichia Pastoris高密度培养条件的摇瓶研究</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=洒荣波&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 洒荣波</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=石贵阳&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,石贵阳</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=王正祥&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,王正祥</a> <span> <a href="/journal-cn-9400/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 食品研究与开发 </a> </span> <span> . 2005</span><span>,第002期</span> </span> </div> </li> <li> <div> <b>4. </b><a class="enjiyixqcontent" href="/academic-journal-cn_journal-east-china-university-science-technology_thesis/0201249532443.html">基因工程菌Pichia pastoris高密度发酵表达重组人血清白蛋白</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=郭美锦&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 郭美锦</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=庄英萍&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,庄英萍</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=吴康华&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,吴康华</a> <span> <a href="/journal-cn-4622/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 华东理工大学学报（自然科学版） </a> </span> <span> . 2002</span><span>,第001期</span> </span> </div> </li> <li> <div> <b>5. </b><a class="enjiyixqcontent" href="/academic-journal-cn_journal-east-china-university-science-technology_thesis/0201249533656.html">基因工程菌Pichia pastoris连续培养的生长及抑制动力学</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=吴康华&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 吴康华</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=郭美锦&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,郭美锦</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=庄英萍&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,庄英萍</a> <span> <a href="/journal-cn-4622/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 华东理工大学学报（自然科学版） </a> </span> <span> . 2001</span><span>,第006期</span> </span> </div> </li> <li> <div> <b>6. </b><a class="enjiyixqcontent" href="/academic-degree-domestic_mphd_thesis/020313714687.html">大肠杆菌胆碱脱氢酶基因在毕氏酵母（pichia pastoris）中的表达</a> <b>[A] </b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=王春明&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 王春明</a> <span> . 2004</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/06120106280260.html">巴斯德毕赤酵母(Pichia pastoris) DAS启动子变体</a> <b>[P]</b> . <span> 中国专利： CN102105589A </span> <span> . 2011-06-22</span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/06120115232.html">重组鱼生长激素基因工程酵母菌pichia pastoris</a> <b>[P]</b> . <span> 中国专利： CN1111200C </span> <span> . 2003.06.11</span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130465903114.html">Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase gene</a> <b>[P]</b> . <span> 外国专利： <!-- 欧洲知识产权局专利： --> EP0374913A1 </span> <span> . 1990-06-27</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：巴斯德毕赤酵母甘油醛-3-磷酸脱氢酶基因 </span> </p> </li> <li> <div> <b>4. </b><a class="enjiyixqcontent" href="/patent-detail/06130466052863.html">PICHIA PASTORIS GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE GENE</a> <b>[P]</b> . <span> 外国专利： <!-- --> CA2000101A1 </span> <span> . 1990-06-22</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：Pichia Pastoris GlyceraldeHyde-3-磷酸脱氢酶基因 </span> </p> </li> <li> <div> <b>5. </b><a class="enjiyixqcontent" href="/patent-detail/06130439896343.html">Promoter and terminator glyceraldehyde 3-phosphate dehydrogenase-1 Pichia methanolica.</a> <b>[P]</b> . <span> 外国专利： <!-- --> ES2231217T3 </span> <span> . 2005-05-16</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：启动子和终止子3-磷酸甘油醛磷酸脱氢酶-1甲醇毕赤酵母。 </span> </p> </li> </ul> </div> </div> </div> <div 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