Purification and Characterization of a Novel Laccase from the Edible Mushroom Hericium coralloides

被引:36
|
作者
Zou, Ya-Jie [1 ]
Wang, He-Xiang [2 ]
Ng, Tzi-Bun [3 ]
Huang, Chen-Yang [1 ]
Zhang, Jin-Xia [1 ]
机构
[1] Chinese Acad Agr Sci, Inst Agr Resources & Reg Planning, Beijing 100081, Peoples R China
[2] China Agr Univ, Coll Biol Sci, State Key Lab Agrobiotechnol, Beijing 100193, Peoples R China
[3] Chinese Univ Hong Kong, Fac Med, Dept Biochem, Shatin, Hong Kong, Peoples R China
关键词
laccase; mushroom; Hericium coralloides; purification; FUNGAL LACCASES; TRAMETES-VERSICOLOR; PLEUROTUS-ERYNGII; FRUITING BODIES; OXIDATION; DERIVATIVES; OXIDASE; SYSTEMS; HEAD;
D O I
10.1007/s12275-012-1372-6
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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 degrees 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 mu M, unlike some previously reported mushroom proteins, but showed significant activity toward human immunodeficiency virus-1 (HIV-1) reverse transcriptase with an IC50 of 0.06 mu M.
引用
收藏
页码:72 / 78
页数:7
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