Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing, 100081, P. R. China.
J Microbiol. 2012 Feb;50(1):72-8. doi: 10.1007/s12275-012-1372-6. Epub 2012 Feb 27.
A novel laccase from the edible mushroom Hericium coralloides was purified by ion exchange chromatography on diethylaminoethyl (DEAE) cellulose, carboxymethyl (CM) cellulose, and Q-Sepharose columns followed by fast protein liquid chromatography gel filtration on a Superdex 75 column. Analysis by gel filtration and SDS-PAGE indicated that the protein is a monomer in solution with a molecular mass of 65 kDa. Its N-terminal amino acid sequence was AVGDDTPQLY, which exhibits partial sequence homology to previously isolated laccases. Optimum activity was observed at pH 2.2 and at 40°C. The enzyme showed activity toward a variety of substrates, the most sensitive of which was 2,2'-azinobis [3-ethylbenzothiazolone-6-sulfonic acid] diammonium salt (ABTS). The degradation activity toward substrates was ABTS > N,N-dimethyl-1,4-phenylenediamine > catechol > 2-methylcatechol > pyrogallol. The laccase did not exert any antiproliferative activity against Hep G2 or MCF 7 tumor cell lines at a concentration of 60 μM, unlike some previously reported mushroom proteins, but showed significant activity toward human immunodeficiency virus-1 (HIV-1) reverse transcriptase with an IC(50) of 0.06 μM.
从食用蘑菇珊瑚菌中纯化得到一种新型漆酶，采用 DEAE 纤维素、CM 纤维素和 Q-Sepharose 柱进行离子交换层析，然后在 Superdex 75 柱上进行快速蛋白液相凝胶过滤层析。凝胶过滤和 SDS-PAGE 分析表明，该蛋白在溶液中为单体，分子量为 65 kDa。其 N 端氨基酸序列为 AVGDDTPQLY，与先前分离的漆酶表现出部分序列同源性。最适活性在 pH 2.2 和 40°C 下观察到。该酶对多种底物表现出活性，最敏感的是 2,2'-联氨双[3-乙基苯并噻唑啉-6-磺酸]二铵盐（ABTS）。对底物的降解活性为 ABTS > N,N-二甲基-1,4-苯二胺 > 儿茶酚 > 2-甲基儿茶酚 > 焦儿茶酚。与一些先前报道的蘑菇蛋白不同，漆酶在 60 μM 浓度下对 Hep G2 或 MCF 7 肿瘤细胞系没有任何抗增殖活性，但对人免疫缺陷病毒-1（HIV-1）逆转录酶表现出显著的活性，IC 50 为 0.06 μM。
