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首页> 外文期刊>Biotechnology and Applied Biochemistry >Improving thermostability of ( RR )‐selective amine transaminase from Aspergillus terreusAspergillus terreus through introduction of disulfide bonds
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Improving thermostability of ( RR )‐selective amine transaminase from Aspergillus terreusAspergillus terreus through introduction of disulfide bonds

机译：提高（ r r） - 从 aspergillus terreus 通过引入二硫键的引入曲霉

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Abstract > To improve the thermostability of ( R )‐selective amine transaminase from Aspergillus terreus (AT‐ATA), we used computer software Disulfide by Design and Modelling of Disulfide Bonds in Proteins to identify mutation sites where the disulfide bonds were most likely to form. We obtained three stabilized mutants (N25C‐A28C, R131C‐D134C, M150C‐M280C) from seven candidates by site‐directed mutagenesis. Compared to the wild type, the best two mutants N25C‐A28C and M150C‐M280C showed improved thermal stability with a 3.1‐ and 3.6‐fold increase in half‐life ( t <sub>1/2</sub> ) at 40 °C and a 4.6 and 5.1 °C increase in T <sub>50</sub> 10 . In addition, the combination of mutant R131C‐D134C and M150C‐M280C displayed the largest shift in thermostability with a 4.6‐fold increase in t <sub>1/2</sub> at 40 °C and a 5.5 °C increase in T <sub>50</sub> 10 . Molecular dynamics simulation indicated that mutations of N25C‐A28C and M150C‐M280C lowered the overall root mean square deviation for the overall residues at elevated temperature and consequently increased the protein rigidity. The stabilized mutation of R131C‐D134C was in the region of high mobility and on the protein surface, and the disulfide bond constraints the flexibility of loop 121–136. </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”> <标题类型=“main”>抽象</ title> >要提高来自曲霉（Ata）（AT-ATA）的温度稳定性（ R </ i>） - 选择性胺转氨酶（ATA），通过设计和建模蛋白质中二硫键的计算机软件二硫化物来鉴定突变二硫键最有可能形成二硫键的遗址。通过点定向诱变，我们获得了来自七个候选物的三种稳定突变体（N25C-A28C，R131C-D134C，M150C-M280C）。与野生型相比，最好的两个突变体N25C-A28C和M150C-M280C显示出改善的热稳定性，半衰期增加3.1-和3.6倍（ T </ i> <sub> 1/2 < /亚>）在40℃和4.6和5.1℃增加 t </ i> <sub> 50 </ sub> 10 </ sup>。另外，突变体R131C-D134C和M150C-M280C的组合显示出热稳定性的最大变化，在40℃和40°C的<I> T <Sub> 1/2 </ Sub>中增加4.6倍。 T </ i> <sub> 50 </ sub> 10 </ sup>增加5.5°C。分子动力学模拟表明，N25C-A28C和M150C-M280C的突变降低了升高温度下总体残留物的总根均方偏差，从而增加了蛋白质刚性。 R131C-D134C的稳定化突变在高迁移率和蛋白质表面上，并且二硫键限制了环121-136的柔韧性。 </ 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>2018年第2期</span><b style="margin: 0 2px;">|</b><span>共8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=Xie Dong‐Fang&option=202" target="_blank" rel="nofollow">Xie Dong‐Fang;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Fang Hui&option=202" target="_blank" rel="nofollow">Fang Hui;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Mei Jia‐Qi&option=202" target="_blank" rel="nofollow">Mei Jia‐Qi;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Gong Jin‐Yan&option=202" target="_blank" rel="nofollow">Gong Jin‐Yan;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Wang Hong‐Peng&option=202" target="_blank" rel="nofollow">Wang Hong‐Peng;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Shen Xiu‐Ying&option=202" target="_blank" rel="nofollow">Shen Xiu‐Ying;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Huang Jun&option=202" target="_blank" rel="nofollow">Huang Jun;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Mei Le‐He&option=202" target="_blank" rel="nofollow">Mei Le‐He;</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>School of Biological and Chemical EngineeringZhejiang University of Science and TechnologyHangzhou People's Republic of China;</p> <p>School of Biological and Chemical EngineeringZhejiang University of Science and TechnologyHangzhou People's Republic of China;</p> <p>Department?of?Chemical?EngineeringUniversity?of?UtahSalt?Lake?City ?UT USA;</p> <p>School of Biological and Chemical EngineeringZhejiang University of Science and TechnologyHangzhou People's Republic of China;</p> <p>School of Biological and Chemical EngineeringZhejiang University of Science and TechnologyHangzhou People's Republic of China;</p> <p>School of Biological and Chemical EngineeringZhejiang University of Science and TechnologyHangzhou People's Republic of China;</p> <p>School of Biological and Chemical EngineeringZhejiang University of Science and TechnologyHangzhou People's Republic of China;</p> <p>Department of Biological and Pharmaceutical EngineeringZhejiang UniversityNingbo People's Republic of 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><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=amine transaminase&option=203" rel="nofollow">amine transaminase;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=disulfide bond&option=203" rel="nofollow">disulfide bond;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=root mean square deviation&option=203" rel="nofollow">root mean square deviation;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=molecular dynamics&option=203" rel="nofollow">molecular dynamics;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=thermostability&option=203" rel="nofollow">thermostability;</a> </p> <div class="translation"> 机译：胺转氨酶;二硫键;根均方偏差;分子动力学;热稳定性; </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> </ul> </div> <div class="similarity_details"> <ul > <li> <div> <b>1. </b><a class="enjiyixqcontent" 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