A novel non-blue laccase from Bacillus amyloliquefaciens: Secretory expression and characterization
A novel non-blue laccase from Bacillus amyloliquefaciens: Secretory expression and characterization
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来自解淀粉芽孢杆菌的新型非蓝色漆酶：分泌表达和表征
DOI：
10.1016/j.ijbiomac.2015.02.019
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发表时间：
2015-05-01
影响因子：
8.2
通讯作者：
Lu, Lei
中科院分区：
文献类型：
--
作者：
Chen, Biao;Xu, Wen-Qi;Lu, Lei
漆酶是一种含铜酶，在许多工业和环境应用中具有广阔的应用前景。本文报道了一种新的非蓝色漆酶的克隆、胞外表达和鉴定。重组酶以高活性分泌到培养上清液中。它缺少蓝色漆酶在610 nm处的典型吸收带。然而，电子顺磁共振（EPR）谱证实了紫外光可见光谱中未检测到的1型铜中心的存在。金属含量分析表明，该酶每蛋白分子含有2个铜离子、1个铁离子和1个锌离子，是一种新型的非蓝色漆酶。重组漆酶的最适pH为6.6℃，最适温度为60℃，最适pH为9.0，最适温度为10 d。在NaCl浓度为200 mM时，酶活性被轻微激活。纯化后的漆酶对活性黑色5和靛蓝胭脂红的脱色效率很高，1 h后脱色率达到93%以上。重组解淀粉酵母菌漆酶具有极强的鲁棒性，与大多数真菌漆酶相比，在各种工业应用中具有许多优势。(C) 2015 Elsevier B.V.版权所有
Laccases are copper-containing enzymes which possess a promising potential in many industrial and environmental applications. Here we describe the cloning, extracellular expression and characterization of a novel non-blue laccase from Bacillus amyloliquefaciens in Pichia pastoris. The recombinant enzyme was secreted into the culture supernatant with high activity. It lacks the absorption band at 610 nm typical for blue laccases. However, electron paramagnetic resonance (EPR) spectrum proved the existence of type 1 copper center that was not detectable in the UV-visible spectrum. Metal content analysis revealed that the enzyme contains two copper ions, one iron ion and one zinc ion per protein molecular, suggesting that it is a novel non-blue laccase. The pH and temperature optima of the recombinant laccase were 6.6 and 60 degrees C, respectively, and it was stable at pH 9.0 for 10 days. The enzyme activity was slightly activated by NaCl with concentration up to 200 mM. The purified laccase showed high efficiency in decolorizing reactive black 5 and indigo carmine, achieving more than 93% decolorization after 1 h. The extreme robustness of the recombinant B. amyloliquefaciens laccase offers several advantages over most fungal laccases in various industrial applications. (C) 2015 Elsevier B.V. All rights reserved.