Cloning and functional analysis of a new laccase gene from Trametes sp 48424 which had the high yield of laccase and strong ability for decolorizing different dyes
Cloning and functional analysis of a new laccase gene from Trametes sp 48424 which had the high yield of laccase and strong ability for decolorizing different dyes
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DOI：
10.1016/j.biortech.2010.10.079
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发表时间：
2011-02-01
影响因子：
11.4
通讯作者：
Yang, Yang
中科院分区：
文献类型：
--
作者：
Fan, Fangfang;Zhuo, Rui;Yang, Yang
从一株新的白腐菌Trametes sp.中克隆了漆酶基因lac48424-1及其全长cDNA，并对其进行了特性分析。48424菌株，漆酶产量高，对不同染料的脱色能力强。Lac48424-1全长1563bp，编码一个成熟的漆酶蛋白，由499个氨基酸组成，由21个氨基酸组成的信号肽组成。推导出的LAC48424-1蛋白序列与已知的其他真菌漆酶高度相似，并含有4个典型漆酶蛋白的铜结合保守区。将编码活性漆酶的lac48424-1基因在巴斯德毕赤酵母中成功表达，验证了该基因的功能。结果表明，该酵母转化子产生的重组漆酶对不同染料具有较强的脱色能力。通过自形成接头聚合酶链式反应获得起始密码子上游的5‘侧翼序列。在lac48424-1的启动子区域发现了许多参与转录调控的顺式反应元件。(C)2010爱思唯尔有限公司。保留所有权利。
The laccase gene lac48424-1 and its corresponding full-length cDNA were cloned and characterized from a novel white-rot fungi Trametes sp. 48424 which had the high yield of laccase and strong ability for decolorizing different dyes. The 1563 bp full-length cDNA of lac48424-1 encoded a mature laccase protein containing 499 amino acids preceded by a signal peptide of 21 amino acids. The deduced protein sequence of LAC48424-1 showed high similarity with other known fungal laccases and contained four copper-binding conserved domains of typical laccase protein. The functionality of lac48424-1 gene encoding active laccase was verified by expressing the gene in the yeast Pichia pastoris successfully. It was found that the recombinant laccase produced by the yeast transformant could decolorize different dyes. The 5'-flanking sequence upstream of start codon was obtained by Self-Formed Adaptor PCR. Many putative cis-acting responsive elements involved in the transcriptional regulation were identified in the promoter region of lac48424-1. (C) 2010 Elsevier Ltd. All rights reserved.