菌物学报

作者：刘天海,张强,苗人云,唐杰,张小平,黄忠乾,彭卫红,谭昊

单位：<sup class=\mag-xml-aff-label-sup added-by-sys\>1.</sup> 四川省农业科学院土壤肥料研究所 四川省农业科学院食用菌研究中心 四川 成都 610066,<sup class=\mag-xml-aff-label-sup added-by-sys\>2.</sup> 四川农业大学资源学院微生物学系 四川 成都 611130,<sup class=\mag-xml-aff-label-sup added-by-sys\>3.</sup> 江南大学生物工程学院 江苏 无锡 214062,<sup class=\mag-xml-aff-label-sup added-by-sys\>4.</sup> 农业部西南区域农业微生物资源利用科学观测实验站 四川 成都 610066

关键词：AA1_2家族,多铜氧化酶,异源表达纯化,漆酶活性,亚铁氧化酶活性

基金：四川省创新能力提升工程项目(2016ZYPZ-028),公益性（农业）行业专项(201503137),国家自然科学基金(31901119)

典型的漆酶通常属于辅助活性酶第一家族第一亚族（auxiliary activity family 1 subfamily 1,简称AA1_1家族）,而AA1_2家族的多铜氧化酶通常拥有将二价铁氧化成三价铁的活性,部分AA1_2家族酶蛋白兼具漆酶活性。梯棱羊肚菌全基因组只有一个AA1_2家族酶基因,该基因编码的酶蛋白是否拥有漆酶功能尚未清楚。本研究主要从酶生化特性的角度,结合酶基因的表达规律,对该基因的功能进行初探。对该AA1_2家族基因在梯棱羊肚菌生长发育不同阶段的表达水平进行实时定量PCR检测;将该基因编码序列克隆到表达载体中在大肠杆菌中异源表达,层析获得纯化的酶蛋白,对酶蛋白的生化特性进行了鉴定。发现该AA1_2多铜氧化酶基因在外源营养袋和土壤中的营养菌丝里低表达,在菇原基和子实体中表达较活跃。异源表达获得纯化的酶蛋白分子量约64kDa,表现出亚铁氧化酶（EC 1.16.3.1）与漆酶（EC 1.10.3.2）双重活性。其亚铁氧化酶活性在pH 4最高,漆酶活性在pH 6最高。亚铁氧化酶活性与漆酶活性的最适温度均为30℃左右,在30℃温育16h后仍保留70%以上活性。亚铁氧化酶和漆酶活性受Mn 2+、Hg 2+和Pb 2+抑制。对蛋白质变性剂SDS、尿素的耐受性较强。本研究通过酶学证据证实了梯棱羊肚菌AA1_2家族多铜氧化酶基因编码的酶蛋白具有亚铁氧化酶-漆酶双重活性,系在子囊菌大型真菌中首次发现,为进一步研究铁元素代谢与漆酶活性在羊肚菌子实体形成与发育过程中的作用提供了启示。"/>template{display:none;}.mag-rich-xref-fn { cursor: pointer;}//长视频$(document).ready(function(){if($("#showLongArticleVideo") && $("#showLongArticleVideo").length && $("#showLongArticleVideo").length>0){mag_ajax_update({ele_id:'showLongArticleVideo',url:mag_currentQikanUrl() + "/CN/article/showLongArticleVideo.do?id="+$("#articleId").val()});}});//可视化图谱$(document).ready(function(){if($("#showArticle_knowledgeMap") && $("#showArticle_knowledgeMap").length && $("#showArticle_knowledgeMap").length>0){jQuery.getJSON("/Jwxb/CN/lexeme/existsCnctstInArticle.do?articleID="+$("#articleId").val(), null, function(json){if(json.result){$("#showArticle_knowledgeMap").show();}});}});$(document).ready(function(){var metaDataStr = $.trim($("#article_metaJson").val());if(metaDataStr!=''){eval("window.metaData="+metaDataStr);}if(window.metaData && (!window.metaData.authorNotesCommon_cn && !window.metaData.authorNotesCorresp_cn && window.metaData.authorNotes_cn)){window.metaData.authorNotesCommon_cn = window.metaData.authorNotes_cn;}if(window.metaData && (!window.metaData.authorNotesCommon_en && !window.metaData.authorNotesCorresp_en && window.metaData.authorNotes_en)){window.metaData.authorNotesCommon_en = window.metaData.authorNotes_en;}var _nlmdtdXml = $("#article_nlmdtdXml").val();var magDir = _nlmdtdXml.replace(/\/[^\/]+$/,'');window.metaData.magDir = magDir;new mag_vue({ el: '#metaVue', //dataUrl: mag_currentQikanUrl() + '/EN/article/getRichHtmlJson.do?articleId='+$("#articleId").val(), data:window.metaData, loading:true, doTextFun:function(text){ return doDataJsonText(text); 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来源：2020年第6期

《菌物学报》期刊编辑部

查看菌物学报杂志2020年第6期