Purification and characterization of the laccase secreted by the white rot fungus Perenniporia tephropora and its role in the decolourization of synthetic dyes
Purification and characterization of the laccase secreted by the white rot fungus Perenniporia tephropora and its role in the decolourization of synthetic dyes
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DOI：
10.1111/j.1365-2672.2006.03152.x
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发表时间：
2007-04-01
影响因子：
4
通讯作者：
Sayadi, S.
中科院分区：
文献类型：
--
作者：
Ben Younes, S.;Mechichi, T.;Sayadi, S.
目的：表征白腐真菌 Perenniporia tephropora 的漆酶，并测试其对合成染料脱色的能力。方法和结果：在所采用的培养条件下，P. tephropora 产生一种漆酶同工酶，通过硫酸铵沉淀、尺寸排阻色谱和阴离子交换色谱将其纯化至电泳均质。该蛋白质是单体，分子量为 63 kDa (SDS-PAGE)，等电点为 3.3。 N端氨基酸序列为SIGPVADLVTNANI，与虎香菇漆酶相似度最高(86.6%)。酶的最适 pH 值各不相同并且取决于底物。 2,6-二甲氧基苯酚（DMP）和2,2'-连氮基-二（3-乙基-苯并噻唑啉-6-磺酸）（ABTS）分别为4.0和5.0。在标准测定条件下，该酶对DMP和ABTS的K-m值分别为7.3和0.4 mmol l(-1)。漆酶被 NaN3、EDTA 和对香豆酸盐抑制，但不被 SDS 和 NaBr 抑制。漆酶在 Cu2+、Co2+、Ca2+、Cd2+、Mg2+、Mn2+、Mo2+、Ni2+、Li+ 和 Al3+ 等金属离子存在下稳定。粗酶和纯化的漆酶能够对纺织工业中的染料进行脱色，包括雷马素亮蓝 R、新兰兰蓝和新兰兰粉红。然而，其他几种染料部分脱色或未脱色。在1-羟基苯并三唑作为介体存在下，仅实现了新兰烷黄的脱色，而大多数染料的脱色仅略有改善。 研究的意义和影响：本研究是第一篇关于从白腐真菌 P. tephropora 中纯化和表征漆酶的报告。这种真菌菌株分泌的高水平漆酶及其稳定性表明它可能是环境应用的有用工具。
Aims: To characterize the white rot fungus Perenniporia tephropora with respect to its laccase and to test its ability to decolourize synthetic dyes.Methods and Results: Under the culture conditions utilized, P. tephropora produced one laccase isozyme, which was purified to electrophoretic homogeneity by ammonium sulfate precipitation, size-exclusion chromatography and anion-exchange chromatography. The protein was monomeric with a molecular mass of 63 kDa (SDS-PAGE) and had an isoelectric point of 3.3. The N-terminal amino acid sequence was SIGPVADLTVTNANI and the highest similarity value was found to the laccase from Lentinus tigrinus (86.6%). The optimum pH of the enzyme varied and was substrate dependent. It was 4.0 and 5.0 for 2,6-dimethoxyphenol (DMP) and 2,2'-azino-di(3-ethyl-benzthiazoline-6-sulfonate) (ABTS), respectively. Under standard assay conditions, K-m values of the enzyme were 7.3 and 0.4 mmol l(-1) towards DMP and ABTS, respectively. The laccase was inhibited by NaN3, EDTA and p-coumarate but not by SDS and NaBr. Laccase was stable in the presence of some metal ions such as Cu2+, Co2+, Ca2+, Cd2+, Mg2+, Mn2+, Mo2+, Ni2+, Li+ and Al3+. The crude enzyme as well as the purified laccase was able to decolourize dyes from the textile industries, including remazol brilliant blue R, neolane blue and neolane pink. However, several other dyes were partially or not decolourized. In the presence of 1-hydroxybenzotriazole as mediator, only the decolourization of neolane yellow was achieved, while the decolourization of most of the dyes was just slightly improved.Significance and Impact of the Study: This study is the first report on the purification and the characterization of the laccase from the white rot fungus P. tephropora. The high levels of laccase secreted by this fungal strain as well as its stability suggest that it could be a useful tool for environmental applications.