高产漆酶革耳的分离鉴定、培养条件、液体发酵及偶氮染料脱色  

Isolation,identification,culture conditions,liquid fermentation,and azo dye decolorization of a high laccase-producing Panus strain

作　　者：李小凤 韩鹏[1] 陈青君[2] 赵智靖 徐诗毅 王秋颖 关体坤 张国庆[2] LI Xiaofeng;HAN Peng;CHEN Qingjun;ZHAO Zhijing;XU Shiyi;WANG Qiuying;GUAN Tikun;ZHANG Guoqing(College of Bioscience and Resource Environment,Beijing University of Agriculture,Key Laboratory of Urban Agriculture(North China)of Ministry of Agriculture and Rural Affairs,Beijing 102206,China;College of Plant Science and Technology,Beijing University of Agriculture,Beijing Advanced Innovation Center for Tree Breeding by Molecular Design,Beijing Key Laboratory for Agricultural Application and New Technique,Beijing 102206,China)

机构地区：[1]北京农学院生物与资源环境学院,农业农村部华北都市农业重点实验室,北京102206 [2]北京农学院植物科学技术学院,北京农学院林木分子设计育种高精尖创新中心,农业应用新技术北京市重点实验室,北京102206

出　　处：《应用与环境生物学报》2022年第3期699-704,共6页Chinese Journal of Applied and Environmental Biology

基　　金：北京市科技计划项目(Z181100009818007);现代农业产业技术体系北京市食用菌创新团队项目(BAIC05-2019);北京农学院学位与研究生教育改革与发展项目(2020YJS040)资助。

摘　　要：漆酶能够高效降解木质素、酚类、胺类等难降解化合物,在造纸、纺织、环保等诸多领域具有良好应用前景.以采集自广西十万大山国家森林公园的高产漆酶革耳属真菌为材料,开展分类鉴定、菌丝最适培养条件、液体发酵产漆酶条件以及染料脱色活性研究.结合形态鉴定与ITS鉴定,确定目标菌株F1625为野生革耳(Panus rudis);结合生长速度和长势,菌丝生长最适碳源为葡萄糖,氮源为酵母浸粉,碳氮比为10/1-20/1,生长因子为维生素B_(1)或维生素C,pH为6-8,温度为32℃,该菌株为高温菌株.液体发酵产漆酶研究表明,PD培养基优于菌丝最适培养条件培养基,Cu^(2+)对产漆酶有抑制作用,而愈创木酚能够诱导漆酶表达.PD培养基添加3.20 mmol/L愈创木酚,在144 h时达到最大酶活,为(4.59±0.04)×10^(3)U/L.F1625菌株发酵液对偶氮染料脱色效果显著,24 h对伊文思蓝和铬黑T的脱色率分别为60.83%±1.40%和65.03%±0.67%.本研究表明F1625菌株在产胞外漆酶和染料降解方面具有潜在应用价值.(图7参26)The enzyme laccase can efficiently catalyze the degradation of lignin,phenols,amines,and other refractory compounds and has good application prospects in multiple fields,including papermaking,textiles,and environmental protection.In this study,wild fruiting bodies of a fungus in the genus Panus with high laccaseproducing activity were collected from the Shiwandashan Mountain National Forest Park in Guangxi Province.We performed strain isolation and classification and determined optimal mycelia culture and liquid fermentation conditions for laccase production.The F1625 strain obtained was systematically classified as Panus rudis based on morphological and internal transcribed spacer(ITS)identification methods.On the basis of mycelial growth rate and quality,we established the following optimal mycelial culture conditions:carbon source,glucose;nitrogen source,yeast extract;carbon:nitrogen(C:N)ratio range,10:1–20:1;growth factors,vitamins B_(1) and C;pH range,6–8;and temperature,32℃.The isolated strain therefore appears to be a thermophilic strain.Examination of laccase production in liquid fermentation revealed that the PD medium was better than the optimal culture medium used for mycelial production.Cu^(2+) was found to inhibit laccase production,whereas guaiacol was found to induce laccase expression.Using PD medium supplemented with 3.20 mmol/L guaiacol,we obtained a maximal enzyme activity of(4.59±0.04)×10^(3)U/L at 144 h.The F1625 strain fermentation broth was found to promote a significant decolorization of azo dyes.After a 24 h incubation,decolorization percentages of 60.83%±1.40%and 65.03%±0.67%were recorded for treated Evans blue and chrome black T dyes,respectively.Our findings indicate that the F1625 strain of Panus rudis has potential applications in extracellular laccase production and dye degradation.

关 键 词：革耳 鉴定 培养条件 漆酶活性 染料脱色 

分 类 号：X703[环境科学与工程—环境工程] X17