Trametes hirsuta漆酶的分离纯化及其对活性染料脱色研究  

Isolation and Purification of Laccase from Trametes hirsuta and Its Application in Reactive Dye Decolorization

作　　者：刘飞 李治宏 张仕豪 刘璇 郑晓晴 焦若若 朱友双 LIU Fei;LI Zhi-hong;ZHANG Shi-hao;LIU Xuan;ZHENG Xiao-qing;JIAO Ruo-ruo;ZHU You-shuang(School of Biological Science/Jining Medical University,Rizhao 276800,China)

机构地区：[1]济宁医学院生物科学学院,山东日照276800

出　　处：《山东农业大学学报（自然科学版）》2024年第1期76-83,共8页Journal of Shandong Agricultural University：Natural Science Edition

基　　金：国家自然科学基金(32000194);山东省高等学校青年创新团队(2022KJ102);济宁医学院贺林院士新医学临床转化工作站科研基金(JYHL2019MS12)。

摘　　要：漆酶是一种含铜的多酚氧化酶,在生物检测、工业染料脱色、有机农药降解、纸浆漂白及食品饮料等领域具有广泛的应用价值。本研究使用实验室自主筛选鉴定的漆酶高产菌株粗毛栓菌(Trametes hirsuta),液态发酵后,培养液经硫酸铵分级沉淀、DEAE Sepharose FF阴离子交换层析分离纯化,酶活总得率57.2%,纯化倍数6.0倍,比活力为758.5 U/mg,漆酶的分子量约为50 kDa。利用粗毛栓菌粗酶液分别对结晶紫、溴酚蓝、孔雀石绿、詹姆斯绿B进行脱色,同时研究了染料浓度、脱色温度、pH和NaCl对溴酚蓝和孔雀石绿脱色率的影响。结果表明,溴酚蓝和孔雀石绿浓度分别为40 mg/L和50 mg/L时脱色率较高;在脱色温度为50℃时,漆酶对溴酚蓝和孔雀石绿的脱色率较高,最高脱色率分别为68.51%和83.06%;溴酚蓝在pH 3.5时脱色率最高达到72.61%,而孔雀石绿的脱色率在pH 4.5时最高达到83.49%;NaCl对Trametes hirsuta漆酶催化染料脱色有一定的抑制作用。本研究表明Trametes hirsuta漆酶在染料脱色中具有较大的应用前景,在工业废水的处理中具有良好的应用潜力。Laccase is a copper-containing polyphenol oxidase with a wide range of application,including bio-detection,industrial dye decolorization,organic pesticide degradation,pulp bleaching,and the food and beverage industries.We utilized a high-yield laccase strain of Tramete hirsuta identified in the laboratory.The laccase was separated and purified through ammonium sulfate precipitation and DEAE Sepharose FF anion exchange chromatography.The enzyme activity yield was 57.2%,with a purification fold of 6.0 and a specific activity of 758.5 U/mg.The molecular weight of laccase was about 50 kDa.The crude enzyme solution from Tramete hirsuta was used to decolorize crystal violet,bromophenol blue,malachite green,and Janus green B.The effects of dye concentration,temperature,pH and NaCl on the decolorization rate of bromophenol blue and malachite green were also investigated.The decolorization rates were higher when the dye concentration was 40 mg/L for bromophenol blue and 50 mg/L for malachite green.The decolorization rates of laccase on bromophenol blue and malachite green were 68.51%and 83.06%,respectively,at the temperature of 50℃.Bromophenol blue exhibited the highest decolorization rate of 72.61%at pH 3.5,while malachite green showed the highest decolorization rate of 83.49%at pH 4.5.NaCl had an inhibitory effect on the dye decolorization catalyzed by Trametes hirsuta laccase.Our study showed Trametes hirsuta laccase has a great application potential in dye decolorization and industrial wastewater treatment.

关 键 词：粗毛栓菌 漆酶 分离纯化 染料脱色 

分 类 号：Q939.5[生物学—微生物学]