Xu LiJing, Wang HeXiang, Ng TziBun
State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing, China.
J Biomed Biotechnol. 2012;2012:536725. doi: 10.1155/2012/536725. Epub 2012 Mar 21.
A 59 kDa laccase with inhibitory activity against HIV-1 reverse transcriptase (IC(50) = 2.4 μM) was isolated from the broth of mycelial culture of the mushroom Lentinus tigrinus. The isolation procedure involved ion exchange chromatography on DEAE-cellulose and CM-cellulose, and gel filtration by fast protein liquid chromatography on Superdex 75. The laccase was adsorbed on both types of ion exchangers. About 95-fold purification was achieved with a 25.9% yield of the enzyme. The procedure resulted in a specific enzyme activity of 76.6 U/mg. Its N-terminal amino acid sequence was GIPDLHDLTV, which showed little similarity to other mushroom laccase and other Lentinus tigrinus strain laccase. Its characteristics were different from previously reported laccase of other Lentinus tigrinus strain. Maximal laccase activity was observed at a pH of 4 and at a temperature of 60°C, respectively. This study yielded the information about the potentially exploitable activities of Lentinus tigrinus laccase.
从虎皮香菇菌丝体培养物的发酵液中分离出一种对HIV-1逆转录酶具有抑制活性(IC(50)=2.4μM)的59 kDa漆酶。分离过程包括在DEAE-纤维素和CM-纤维素上进行离子交换色谱,以及在Superdex 75上通过快速蛋白质液相色谱进行凝胶过滤。该漆酶能吸附在两种类型的离子交换剂上。酶的产量为25.9%,实现了约95倍的纯化。该方法得到的酶比活性为76.6 U/mg。其N端氨基酸序列为GIPDLHDLTV,与其他香菇漆酶和其他虎皮香菇菌株漆酶几乎没有相似性。其特性与先前报道的其他虎皮香菇菌株的漆酶不同。漆酶的最大活性分别在pH值为4和温度为60°C时观察到。本研究获得了有关虎皮香菇漆酶潜在可利用活性的信息。