Optimization of defined medium for recombinant Komagataella phaffii expressing cyclodextrin glycosyltransferase

Zhang Jianguo; Zhao Yixin; Li Mengla; Liu Taiyu

摘要

Abstract > The cyclodextrin glycosyltransferase (CGTase) is an important enzyme for cyclodextrin (CD) production, and is also widely used in the biotechnology, food, and pharmaceuticals industries. Secretory CGTase production by recombinant Komagataella phaffii using defined medium is a promising approach because of low cost, less impurity protein. It was found that no CGTase was expressed using traditional defined medium (basal salt medium [BSM]) because of pH value decreasing significantly. CGTase was expressed by recombinant K. phaffii through pH maintenance in range of 5.5–7.0. β‐CGTase activity increased to 122.0 U/mL after optimization of glycerol, phosphate buffer, pH value, ammonium sulfate, temperature, methanol, and additives based on BSM, establishing a modified defined medium. These results showed that it was necessary to establish recombinant K. phaffii ‐based special defined medium although the same host cell used for different heterologous protein expression. </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> > 环糊精糖基转移酶（CGTase）是环糊精（CD）生产的重要酶，也广泛应用于生物技术，食品和制药行业。分泌CGTase通过重组产生 Komagataella phaffii </ i> 使用定义的培养基是一种有希望的方法，因为成本低，杂质蛋白质较少。发现由于pH值显着降低，使用传统的定义培养基（基础盐介质[BSM]）表示没有CGTase。 CGTase由重组表达 k。 phaffii </ i> 通过pH维护范围为5.5-7.0。在优化甘油，磷酸盐缓冲液，pH值，硫酸铵，温度，甲醇和基于BSM的添加剂后，β-CgTase活性增加至122.0u / ml，建立改性定义的培养基。这些结果表明有必要建立重组 k。 phaffii </ i> 基于特殊定义的培养基，尽管用于不同异源蛋白表达的相同宿主细胞。 </ 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>2019年第5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=Zhang Jianguo&option=202" target="_blank" rel="nofollow">Zhang Jianguo;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Zhao Yixin&option=202" target="_blank" rel="nofollow">Zhao Yixin;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Li Mengla&option=202" target="_blank" rel="nofollow">Li Mengla;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Liu Taiyu&option=202" target="_blank" rel="nofollow">Liu Taiyu;</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>Institute of Food Science and Engineering School of Medical Instrument and Food Engineering University of Shanghai for Science and TechnologyShanghai China;</p> <p>Institute of Food Science and Engineering School of Medical Instrument and Food Engineering University of Shanghai for Science and TechnologyShanghai China;</p> <p>Institute of Food Science and Engineering School of Medical Instrument and Food Engineering University of Shanghai for Science and TechnologyShanghai China;</p> <p>Institute of Food Science and Engineering School of Medical Instrument and Food Engineering University of Shanghai for Science and TechnologyShanghai 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=cyclodextrin glycosyltransferase&option=203" rel="nofollow">cyclodextrin glycosyltransferase;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=defined medium&option=203" rel="nofollow">defined medium;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Komagataella phaffii&option=203" rel="nofollow">Komagataella phaffii;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=optimization&option=203" rel="nofollow">optimization;</a> <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> </p> <div class="translation"> 机译：环糊精糖基转移酶;定义的培养基;Komagataella phaffii;优化;Pichia Pastoris; </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="/academic-journal-cn_chinese-journal-oceanic-limnology-english-edition_thesis/0201252506907.html">Statistical optimization of medium composition and culture condition for the production of recombinant antilipopolysaccharide 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Muhammad Tariq Navid</a> <span> . 2016</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/06130403979667.html">KOMAGATAELLA PHAFFII RECOMBINANT XYLIC ACID PRODUCER AND METHOD FOR THE PRODUCTION OF XYLIC ACID FROM XYLOSIS</a> <b>[P]</b> . <span> 外国专利：  BR102018001359A2 </span> <span> . 2019-08-06</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：KOMAGATAELLA PHAFFII重组木糖生产者和从木糖虫生产木糖的方法 </span> </p> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/06130499831146.html">METHOD FOR PRODUCING PHOSPHOLIPASE А2 KOMAGATAELLA PHAFFII (PICHIA PASTORIS) YIB ΔLEU2_PLA2S PRODUCTION STRAIN</a> <b>[P]</b> . <span> 外国专利：  RU2746817C1 </span> <span> . 2021-04-21</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：生产磷脂酶的方法а2Komagataella phaffii（Pichia pastoris）YibΔLeu2_PLA2S生产菌株 </span> </p> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130500717350.html">KOMAGATAELLA PHAFFII YEAST TRANSFORMANT THAT PRODUCES CRONOBACTER TURICENSIS PHYTASE</a> <b>[P]</b> . <span> 外国专利：  RU2756330C1 </span> <span> . 2021-09-29</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：Komagataella phaffii酵母转化体，产生植物植物植酸酶 </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 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