一种白腐真菌的分离、鉴定、培养及产漆酶条件  

Isolation, identification, culture conditions, and laccase production of white rot fungus

作　　者：胡渤洋 王寿南 陈青君[2] 张国庆 杨佳玥 徐鑫 韩鹏 张宇轩 李兵 HU Boyang;WANG Shounan;CHEN Qingjun;ZHANG Guoqing;YANG Jiayue;XU Xin;HAN Peng;ZHANG Yuxuan;LI Bing(College of Biological Science and Engineering, Beijing University of Agriculture, Key Laboratory of Urban Agriculture （North China） of Ministry of Agriculture, Beijing 102206, China;Plant Science and Technology College, Beijing University of Agriculture, Beijing 102206, China)

机构地区：[1]北京农学院生物科学与工程学院、农业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,北京102206

出　　处：《应用与环境生物学报》2018年第2期367-373,共7页Chinese Journal of Applied and Environmental Biology

基　　金：北京市自然科学基金项目(5162006);北京市科技新星计划项目(xx2015B025);农业部华北都市农业重点实验室开放课题(kf 2017012);北京市本科生科学研究计划项目资助~~

摘　　要：白腐真菌是一类降解木质素使木材形成白色腐朽的担子菌，往往具有高产胞外漆酶的活性. 从北京农学院校园内分离到一株高温型、高产漆酶白腐真菌菌株，编号为BUA-01. 结合形态学特征与ITS序列分析，确定其分类学地位；进一步研究其菌丝生长的最适碳源、氮源、碳氮比（C/N）、生长因子、最适温度和最适pH值；利用不同浓度Cu^2＋（CuSO4）的诱导，探讨液体发酵条件下对胞外漆酶产量的影响；通过向发酵液中加入3种偶氮染料（依文思蓝、甲基橙和铬黑T），研究菌株对染料的降解效果. 结果表明，该菌株与毛栓菌（Trametes hirsuta）的同源性最高，为100%，遗传距离为0，确定BUA-01菌株为栓菌属（Trametes）真菌. 菌丝体生长的最适碳源为淀粉，最适氮源为黄豆粉、酵母浸粉，最适C/N值为40/1和10/1，最适温度为37 ℃，最适pH为6.0-7.0，供试生长因子对菌丝生长无显著促进作用. 0.25 mmol/L的Cu^2＋对胞外漆酶产量有极显著的促进作用，在96 h时，发酵液的活性达到最高，为1081.33 ± 6.3 U/mL，是对照组的26倍. BUA-01菌株对偶氮染料降解效果显著，12 h对依文思蓝、甲基橙和铬黑T的脱色率分别为93.31% ± 0.16%、92.37% ± 0.42%和79.25% ± 0.64%. 本研究表明菌株BUA-01在产胞外漆酶和染料降解方面具有潜在应用价值. （图3 表7 参42）The white rot fungi are members of Basidiomycota, which can degrade lignin and form white rot. They are high producers of extracellular laccases. In the present study, pure culture strain of high-temperature and high-laccase production types （numbered as BUA-01） was isolated from the fruiting bodies of a white rot fungus collected in the campus of Beijing University of Agriculture. The taxonomic characteristic was determined based on morphological and ITS sequence analysis. Furthermore, the optimal culture conditions for the mycelia were determined, including carbon source, nitrogen source, C/N ratio, growth factors, temperature, and pH. Extracellular laccase production was investigated in liquid fermentation with different concentrations of Cu^2＋ （CuSO4） as inducer. Decolorizing activity of the fermentation broth was assayed using three azo dyes： Evans blue, methyl orange, and eriochrome black T. The results showed that the strain possessed the highest homology toward Trametes hirsuta, with the homology ratio of 100% and the genetic distance of 0, suggesting that the strain BUA-01 belonged to the genus Trametes. The culture condition investigated revealed that the optimal condition for mycelia growth included the following： carbon source, starch; nitrogen source, soybean powder and yeast extract; C/N ratio, 40/1 and 10/1; temperature, 37 ℃; and pH, 6.0-7.0. The assayed growth factors had no significant effect on mycelial growth. It demonstrated high laccase activity in liquid fermentation. The highest extracellular laccase activity of 1 081.33 ± 6.3 U/mL was observed in the broth with a Cu^2＋ adjunction concentration of 0.25 mmol/L after a 96-h culture period. It was about 26-fold higher than that of the control group. The isolated strain exhibited significant decolorizing activity toward the azo dyes Evans blue, methyl orange, and eriochrome black T, with the decolorization rate at 12 h of 93.31% ± 0.16%, 92.37% ± 0.42%, 79.25% ± 0.64%, respectively. This suggests that the strain possesses

关 键 词：栓菌 鉴定 培养条件 漆酶 染料降解 

分 类 号：Q939.99[生物学—微生物学] X172[环境科学与工程—环境科学]