Production of Hydroxynitrile Lyase from        <i >Davallia tyermanniiDavallia tyermannii        (        <i >DtDt        HNL) in        <i >Komagataella phaffiiKomagataella phaffii        and Its Immobilization as a CLEA to Generate a Robust Biocatalyst

Lanfranchi Elisa; Grill Birgit; Raghoebar Zainab; Van?Pelt Sander; Sheldon Roger A.; Steiner Kerstin; Glieder Anton; Winkler Margit

摘要

Abstract > Hydroxynitrile lyase from the white rabbit's foot fern Davallia tyermannii ( Dt HNL) catalyzes the enantioselective synthesis of α‐cyanohydrins, which are key building blocks for pharmaceutical and agrochemical industries. An efficient and competitive process necessitates the availability and robustness of the biocatalyst. Herein, the recombinant production of Dt HNL1 in Komagataella phaffii , yielding approximately 900?000?U?L ?1 , is described. Dt HNL1 constitutes approximately 80?% of the total protein content. The crude enzyme was immobilized. Crosslinked enzyme aggregates (CLEAs) resulted in significant enhancement of the biocatalyst stability under acidic conditions (activity retained after 168?h at pH?2.4). The Dt HNL1–CLEA was employed for ( R )‐mandelonitrile synthesis (99?% conversion, 98?% enantiomeric excess) in a biphasic system, and evaluated for the synthesis of ( R )‐hydroxypivaldehyde cyanohydrin under reaction conditions that immediately inactivated non‐immobilized Dt HNL1. The results show the Dt HNL1–CLEA to be a stable biocatalyst for the synthesis of enantiomerically pure cyanohydrins under acidic conditions. </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> >羟基硝酶从白色兔子的脚蕨 davallia tyermannii </ i>（< I> DT </ i> HNL）催化α-氰基氢胺的对映选择性合成，这是药品和农用化学行业的关键构建块。有效且竞争的过程需要生物催化剂的可用性和稳健性。在此，描述了在 Komagataella phaffii中的重组产生 komagatella phaffii </ i>，得到约900Ω·000？uΔ1Δ1</ sup>。 DT </ i> HNL1构成总蛋白质含量的约80倍。固定粗酶。交联的酶聚集体（CLEA）导致在酸性条件下的生物催化剂稳定性的显着提高（在pH？2.4的168℃后保留的活性。使用 DT </ i> HNL1-CLEA用于（ R） - 摩尔腈合成（99〜％转化率，98〜％对映体过量），并评估合成在反应条件下（ R </ i>） - 羟基戊二醛氰基醇在立即灭活未固定的 DT </ i> HNL1。结果表明， DT </ i> HNL1-CLEA是稳定的生物催化剂，用于在酸性条件下合成对映体纯氰醇。 </ 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4期</span><b style="margin: 0 2px;">|</b><span>共5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=Lanfranchi Elisa&option=202" target="_blank" rel="nofollow">Lanfranchi Elisa;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Grill Birgit&option=202" target="_blank" rel="nofollow">Grill Birgit;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Raghoebar Zainab&option=202" target="_blank" rel="nofollow">Raghoebar Zainab;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Van?Pelt Sander&option=202" target="_blank" rel="nofollow">Van?Pelt Sander;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Sheldon Roger A.&option=202" target="_blank" rel="nofollow">Sheldon Roger A.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Steiner Kerstin&option=202" target="_blank" rel="nofollow">Steiner Kerstin;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Glieder Anton&option=202" target="_blank" rel="nofollow">Glieder Anton;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Winkler Margit&option=202" target="_blank" rel="nofollow">Winkler Margit;</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>acib GmbHPetersgasse 14 8010 Graz Austria;</p> <p>acib GmbHPetersgasse 14 8010 Graz Austria;</p> <p>CLEA-TechnologiesDelftechpark 34 2628 XH Delft The Netherlands;</p> <p>CLEA-TechnologiesDelftechpark 34 2628 XH Delft The Netherlands;</p> <p>Molecular Sciences InstituteUniversity of the WitwatersrandJohannesburg PO Wits 2050 South Africa;</p> <p>acib GmbHPetersgasse 14 8010 Graz Austria;</p> <p>acib GmbHPetersgasse 14 8010 Graz Austria;</p> <p>acib GmbHPetersgasse 14 8010 Graz Austria;</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=crosslinked enzyme aggregates&option=203" rel="nofollow">crosslinked enzyme aggregates;</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=hydroxynitrile lyase&option=203" rel="nofollow">hydroxynitrile lyase;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=immobilization&option=203" rel="nofollow">immobilization;</a> </p> <div class="translation"> 机译：生物催化;交联酶聚集体;酶;羟腈裂解酶;固定化; 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