Decolorization of different dyes by a newly isolated white-rot fungi strain Ganoderma sp.En3 and cloning and functional analysis of its laccase gene.
Decolorization of different dyes by a newly isolated white-rot fungi strain Ganoderma sp.En3 and cloning and functional analysis of its laccase gene.
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DOI：
10.1016/j.jhazmat.2011.05.106
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发表时间：
2011-08
影响因子：
13.6
通讯作者：
Rui Zhuo;Li Ma;Fangfang Fan;Yangmin Gong;Xia Wan;Mulan Jiang;Xiaoyu Zhang;Yang Yang-Yang
中科院分区：
文献类型：
--
作者：
Rui Zhuo;Li Ma;Fangfang Fan;Yangmin Gong;Xia Wan;Mulan Jiang;Xiaoyu Zhang;Yang Yang-Yang
从中国紫金山林区新分离到一株产漆酶的白腐真菌灵芝（Ganoderma sp. En 3）。灵芝菌En 3对4种合成染料、2种模拟染浴废水和真实的印染废水均有较强的脱色能力。在脱色过程中对漆酶活性的诱导表明，漆酶在该菌对不同染料的高效脱色过程中起着重要作用。对小麦和水稻的毒性研究表明，灵芝En 3菌株对4种合成染料、2种模拟染料废水和真实的纺织染料废水具有良好的解毒作用。克隆了灵芝漆酶基因lac-En 3 -1及其相应的全长cDNA。LAC-En 3 -1的蛋白质序列含有4个典型漆酶蛋白铜结合保守结构域。通过在毕赤酵母中成功表达lac-En 3 -1基因，验证了lac-En 3 -1基因的功能。该菌株所产的重组漆酶对合成染料、模拟染料废水和真实的印染废水均有脱色作用。漆酶对不同染料的脱色能力与漆酶活性呈正相关。此外，还获得了lac-En 3 -1基因起始密码子ATG上游的5′侧翼序列。在lac-En 3 -1的启动子区域预测了许多推定的顺式作用响应元件。
A laccase-producing white-rot fungi strain Ganoderma sp.En3 was newly isolated from the forest of Tzu-chin Mountain in China. Ganoderma sp.En3 had a strong ability of decolorizing four synthetic dyes, two simulated dye bath effluents and the real textile dye effluent. Induction in the activity of laccase during the decolorization process indicated that laccase played an important role in the efficient decolorization of different dyes by this fungus. Phytotoxicity study with respect to Triticum aestivum and Oryza sativa demonstrated that Ganoderma sp.En3 was able to detoxify four synthetic dyes, two simulated dye effluents and the real textile dye effluent. The laccase gene lac-En3-1 and its corresponding full-length cDNA were then cloned and characterized from Ganoderma sp.En3. The deduced protein sequence of LAC-En3-1 contained four copper-binding conserved domains of typical laccase protein. The functionality of lac-En3-1 gene encoding active laccase was verified by expressing this gene in the yeast Pichia pastoris successfully. The recombinant laccase produced by the yeast transformant could decolorize the synthetic dyes, simulated dye effluents and the real textile dye effluent. The ability of decolorizing different dyes was positively related to the laccase activity. In addition, the 5′-flanking sequence upstream of the start codon ATG in lac-En3-1 gene was obtained. Many putative cis-acting responsive elements were predicted in the promoter region of lac-En3-1.