Redesign of a New Manganese Peroxidase Highly Expressed in        <i >Pichia pastorisPichia pastoris        towards a Lignin‐Degrading Versatile Peroxidase

Bronikowski Agathe; Koschorreck Katja; Urlacher Vlada B.

摘要

Abstract > Manganese peroxidases, lignin peroxidases, and versatile peroxidases secreted by white rot fungi are supposed to play an essential role in lignin degradation. Thus, these enzymes have attracted significant attention as potential biocatalysts. Versatile peroxidases are the most interesting ones, since they comprise activities of manganese and lignin peroxidases. Herein, we demonstrate how the properties of a new manganese peroxidase from Moniliophthora roreri , designated MrMnP1, were shifted towards those of a versatile peroxidase. MrMnP1 was cloned in Pichia pastoris X‐33 and highly expressed in a fed‐batch fermentation, yielding 132?mg?L ?1 of active enzyme. To extend the substrate spectrum of MrMnP1, a catalytically active tryptophan present in lignin and versatile peroxidases was first introduced. Additionally, the role of five amino acids at positions adjacent to the catalytic tryptophan was elucidated through their replacement by those found in a versatile peroxidase from Pleurotus eryngii . The resulting mutants demonstrated new activities towards high‐redox‐potential substrates, such as lignin dimers, veratryl alcohol, and the azo dye Reactive Black?5. </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”>摘要</标题> >锰过氧化物酶，木质素过氧化物酶和白腐真菌分泌的通酯过氧化物酶应该发挥作用木质素劣化中的基本作用。因此，这些酶引起了潜在的生物催化剂的显着关注。多功能过氧化物酶是最有趣的酶，因为它们包括锰和木质素过氧化物酶的活性。在此，我们证明了如何从单胞嘧啶roreri </ i>指定MRMNP1的新锰过氧化物酶的性质转移到通用过氧化物酶的那些。 MRMNP1在 Pichia Pastoris X / I> X-33中克隆并在FED批量发酵中高度表达，得到132μm≤Xα1的活性酶。为了延长MRMNP1的基材光谱，首先引入木质素中存在的催化活性色氨酸和过氧化物酶。另外，通过替代在来自 Pleurotus erryngii的替代物中替代，阐明了五个氨基酸在邻近催化色氨酸的位置的作用。所得突变体展示了对高氧化还原潜在底物的新活动，例如木质素二聚体，维拉醇和偶氮染料反应性黑色α5。 </ 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-16039/'>《Chembiochem: A European journal of chemical biology》</a> <b style="margin: 0 2px;">|</b><span>2018年第23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=Bronikowski Agathe&option=202" target="_blank" rel="nofollow">Bronikowski Agathe;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Koschorreck Katja&option=202" target="_blank" rel="nofollow">Koschorreck Katja;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Urlacher Vlada B.&option=202" target="_blank" rel="nofollow">Urlacher Vlada B.;</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 Biochemistry and Bioeconomy Science Center (BioSc)Heinrich Heine University DüsseldorfUniversit?tsstrasse 1 40225 Düsseldorf Germany;</p> <p>Institute of Biochemistry and Bioeconomy Science Center (BioSc)Heinrich Heine University DüsseldorfUniversit?tsstrasse 1 40225 Düsseldorf Germany;</p> <p>Institute of Biochemistry and Bioeconomy Science Center (BioSc)Heinrich Heine University DüsseldorfUniversit?tsstrasse 1 40225 Düsseldorf 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/179.html" title="分子生物学">分子生物学;</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=biocatalysis&option=203" rel="nofollow">biocatalysis;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=enzymes&option=203" rel="nofollow">enzymes;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=mutagenesis&option=203" rel="nofollow">mutagenesis;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=renewable resources&option=203" rel="nofollow">renewable resources;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=structure–activity relationships&option=203" rel="nofollow">structure–activity relationships;</a> </p> <div class="translation"> 机译：生物催化;酶;诱变;可再生资源;结构 - 活动关系; </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_detail_thesis/0201281158544.html">Enhanced Palladium Adsorption by Immobilized Pichia pastoris: A Reusable Catalyst for Reduction of 4-Nitrophenol</a> <b>[J]</b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Liqin Lin&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Liqin Lin,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Xiaolian Jing&option=202" target="_blank" rel="nofollow" class="tuijian_auth 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REDDY,</a> <a href="/conference-foreign-190587/" target="_blank" rel="nofollow" class="tuijian_authcolor">Biotechnology for Fuels and Chemicals .</a> <span>1998</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：木质素过氧化物酶和锰过氧化物酶体外降解水性介质中的不溶性木质素</span> </p> </li> <li> <div> <b>6. </b><a class="enjiyixqcontent" href="/academic-degree-foreign_mphd_thesis/02061721820.html">Expression of recombinant manganese peroxidase in Pichia pastoris: Biosynthesis, stability and kinetics analysis</a> <b>[D] </b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Jiang, Fei&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Jiang, Fei </a> <span>2006</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：重组锰过氧化物酶在毕赤酵母中的表达：生物合成，稳定性和动力学分析</span> </p> </li> <li> <div> <b>7. </b><a class="enjiyixqcontent" href="/academic-journal-foreign-pmc_detail_thesis/040003365928.html">Expression and characteristics of manganese peroxidase from Ganoderma lucidum in Pichia pastoris and its application in the degradation of four dyes and phenol</a> <b>[O] </b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Hui Xu&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Hui Xu,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Meng-Yuan Guo&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Meng-Yuan Guo,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Yan-Hua Gao&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">Yan-Hua Gao,</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：灵芝中锰过氧化物酶在毕赤酵母中的表达特性及其在降解四种染料和苯酚中的应用</span> </p> </li> <li> <div> <b>8. </b><a class="enjiyixqcontent" href="/open-access_resources_thesis/0100020955116.html ">Ligninolytic properties of highly expressed transformant with novel lignin peroxidase from hyper lignin-degrading fungus Phanerochaete sordida YK-624</a> <b>[O] </b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=杉浦, 立樹&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">杉浦, 立樹,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=山岸, 賢治&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">山岸, 賢治,</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=平井, 浩文&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">平井, 浩文,</a> <span>2010</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：木质素过度降解真菌Phanerochaete sordida YK-624新型木质素过氧化物酶高表达转化子的木质素分解特性</span> </p> </li> <li> <div> <b>9. </b><a class="enjiyixqcontent" href="/ntis-science-report_pb_thesis/020711041654.html">Lignin Degradation and Lignin Peroxidase Production in Cultures of 'Phanerochaetechrysosporium' Immobilized on Porous Ceramic Supports</a> <b>[R] </b> . <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Cornwell, K. 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J.,</a> <span>1990</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：多孔陶瓷载体固定化'phanerochaetechrysosporium'培养物中木质素降解和木质素过氧化物酶的产生</span> </p> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/academic-journal-cn_detail_thesis/0201292474561.html">在甲醇酵母Pichia pastoris胞内表达有活性的辣根过氧化物酶</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=蒋太交&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 蒋太交</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=吉鑫松&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,吉鑫松</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=章如安&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,章如安</a> <span> <a href="/journal-cn-56721/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 中国生物化学与分子生物学报 </a> </span> <span> . 2000</span><span>,第2期</span> </span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/academic-journal-cn_progress-biochemistry-biophysics_thesis/0201293658967.html">甲醇酵母Pichia pastoris高水平表达有活性的辣根过氧化物酶</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=蒋太交&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 蒋太交</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=吉鑫松&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,吉鑫松</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=章如安&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,章如安</a> <span> <a href="/journal-cn-5079/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 生物化学与生物物理进展 </a> </span> <span> . 1999</span><span>,第6期</span> </span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/academic-journal-cn_fudan-journal-natural-science-edition_thesis/0201297771803.html">一个新的PMA诱导相关基因在Pichia pastoris酵母系统中的表达</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=黄燕&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 黄燕</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=吴超群&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,吴超群</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=和桂芬&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,和桂芬</a> <span> <a href="/journal-cn-50345/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 复旦学报：自然科学版 </a> </span> <span> . 2000</span><span>,第6期</span> </span> </div> </li> <li> <div> <b>4. </b><a class="enjiyixqcontent" href="/academic-journal-cn_journal-hebei-agricultural-university_thesis/0201281863169.html">谷氨酰胺转氨酶在Pichia pastoris GS115中的表达及其发酵条件响应面法优化</a> <b>[J]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=于凡&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 于凡</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=刘双&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,刘双</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=陈海清&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,陈海清</a> <span> <a href="/journal-cn-11519/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 河北农业大学学报 </a> </span> <span> . 2020</span><span>,第006期</span> </span> </div> </li> <li> <div> <b>5. </b><a class="enjiyixqcontent" href="/academic-conference-cn_meeting-9664_thesis/020222341931.html">Effective and stable porcine interferon-a production by Pichia pastoris fed-batch cultivation with multi-variables clustering and analysis</a> <b>[C]</b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ruisong Yu&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . Ruisong Yu</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Shijuan Dong&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Shijuan Dong</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Yumin Zhu&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor">,Yumin Zhu</a> <span> <a href="/conference-cn-9664/" target="_blank" rel="nofollow" class="tuijian_authcolor"> . 上海市畜牧兽医学会2011年学术年会 </a> <span> <span> . 2011</span> </span> </div> </li> <li> <div> <b>6. </b><a class="enjiyixqcontent" href="/academic-degree-domestic_mphd_thesis/020316245991.html">谷氨酰胺转氨酶在Pichia pastoris GS115中的表达及其发酵条件优化</a> <b>[A] </b> <span> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=于凡&option=202" target="_blank" rel="nofollow" class="tuijian_auth tuijian_authcolor"> . 于凡</a> <span> . 2020</span> </span> </div> </li> </ul> <ul style="display: none;"> <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/patent-detail/06120103863246.html">一种定量Pichia pastoris细胞残留的核酸检测试剂盒</a> <b>[P]</b> . <span> 中国专利： CN111334607A </span> <span> . 2020-06-26</span> </div> </li> <li> <div> <b>2. </b><a class="enjiyixqcontent" href="/patent-detail/061201614456.html">在Pichia pastoris中组成型表达菌丝霉素衍生物MP1102的方法</a> <b>[P]</b> . <span> 中国专利： CN103333912B </span> <span> . 2015.04.15</span> </div> </li> <li> <div> <b>3. </b><a class="enjiyixqcontent" href="/patent-detail/06130418567585.html">Oligonucleotide for detecting lignin peroxidase gene and manganese peroxidase gene, and screening method for lignin peroxidase gene and manganese peroxidase gene using the same</a> <b>[P]</b> . <span> 外国专利：  JP5567360B2 </span> <span> . 2014-08-06</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：用于检测木质素过氧化物酶基因和锰过氧化物酶基因的寡核苷酸，以及使用该寡核苷酸的木质素过氧化物酶基因和锰过氧化物酶基因的筛选方法 </span> </p> </li> <li> <div> <b>4. </b><a class="enjiyixqcontent" href="/patent-detail/06130426581684.html">OLIGONUCLEOTIDE FOR DETECTING LIGNIN PEROXIDASE GENE AND MANGANESE PEROXIDASE GENE, AND METHOD FOR SCREENING LIGNIN PEROXIDASE GENE AND MANGANESE PEROXIDASE GENE USING THE SAME</a> <b>[P]</b> . <span> 外国专利：  JP2011160770A </span> <span> . 2011-08-25</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：用于检测木质素过氧化物酶基因和锰过氧化物酶基因的寡核苷酸，以及使用相同方法筛选木质素过氧化物酶基因和锰过氧化物酶基因的方法 </span> </p> </li> <li> <div> <b>5. </b><a class="enjiyixqcontent" href="/patent-detail/06130451459229.html">Phanerochaete chrysospoirium strains CNCM I-1511, I-1512 and I- 1513 for producing lignin peroxidase and manganese peroxidase</a> <b>[P]</b> . <span> 外国专利：  US5972672A </span> <span> . 1999-10-26</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：Phanerochaete chrysospoirium菌株CNCM I-1511，I-1512和I-1513用于生产木质素过氧化物酶和锰过氧化物酶 </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 id="thesis_get_original1" class="downloadBth" style="bottom: 19px;z-index: 999;" onclick="ywcd('0704022056533','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="关闭" 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 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Redesign of a New Manganese Peroxidase Highly Expressed in Pichia pastorisPichia pastoris towards a Lignin‐Degrading Versatile Peroxidase

摘要