产漆酶疣孢漆斑菌NF-05的分离及对偶氮染料的脱色  

Isolation and decolorization of azo dye of a laccase-producing Myrothecium verrucaria NF-05

作　　者：赵敏[1] 王海东[1] 赵丹[1,2] 谷惠琦[1] 张曦[1] 

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040 [2]黑龙江大学生命科学学院微生物省高校重点实验室,哈尔滨150080

出　　处：《菌物学报》2011年第4期604-611,共8页Mycosystema

基　　金：世界自然基金(No.WWF;CN0078.01);国家自然科学基金(No.30170775;30671702);国家支撑项目(No.2008BADA1B01-10)

摘　　要：以木质素磺酸钠为唯一碳源的培养基对带岭凉水自然保护区土壤样品进行富集培养,涂布于愈创木酚-PDA平板。经2,2′-连氮-双(3-乙基苯并噻唑-6-磺酸)(ABTS)和丁香醛联氮(SGZ)平板检测初筛,ABTS法测定摇瓶发酵液酶活力复筛,筛选到一株漆酶高产真菌NF-05。形态学观察结合rDNA-ITS序列分析,鉴定该菌为半知菌疣孢漆斑菌Myrothecium verrucaria。该菌株在液体产酶培养基中生物量积累与产酶基本同步,发酵第5天达到产酶高峰,最高酶活力为8,375.87U/L。纯化漆酶对偶氮染料脱色研究结果表明,该酶在96h对甲基橙脱色率达到90%以上,以2,2,6,6-四甲基哌啶氧化物(TE)为介体时,48h脱色率即达90%以上;该酶在24h对橙黄Ⅰ的脱色率即达90%以上;以TE为介体时,该酶在24h即使橙黄G6完全脱色。Soil samples were collected in Liangshui Nature Reserve,China and enriched in medium with lignin-sodium salfacid as sole carbon source.Guaiacol-PDA plates were used to separate laccase-producing fungi.A high-yield strain NF-05 was screened after testing with 2,2′-azino-bis（3-ethylbenz-thiazoline-6-sulfonic acid） （ABTS） and 4-hy-droxy-3,5-dimethoxybenzaldehyde azine （SGZ）.NF-05 was identified as Myrothecium verrucaria based on morphological characteristics and sequence analysis of rDNA-ITS.Laccase production of NF-05 showed synchronism in concordance with hypha growth in fermentation medium.Enzyme activity reached the peak of 8,375.87U/L at the fifth day.Laccase from NF-05 was purified for assay of decolorizing azo dyes with mediators,which resulted in a decolorization percentage of 90% for methyl orange mediated by tetramethylpiperidinooxy （TE）,90.1% for orangeⅠand 100% for orange G6 mediated by TE.

关 键 词：漆酶 半知菌 疣孢漆斑菌 偶氮染料 介体 

分 类 号：TQ925[轻工技术与工程—发酵工程]