Purification and Characterization of An Extracellular Laccase with Antiproliferative Activity from The Sanghuang Mushroom Inonotus baumii
We described the purification and characterization of a novel extracellular laccase from the traditional Chinese medicinal mushroom Inonotus baumii with antiproliferative activity.The laccase (IBL) was purified from fermentation broth of I.baumii by employing initial filtration and centrifugation steps,followed by three ion-exchange chromatography steps comprising DEAE-cellulose,CM-cellulose,and Q-Sepharose,and a final gel-filtration step by fast protein liquid chromatography (FPLC) on Superdex 75.The purified enzyme was a monomeric protein with a molecular mass of 66 kDa calculated by FPLC and SDS-PAGE.It possessed an N-terminal amino acid sequence of AIGPVDEV (SPIN:C0HJB2),a temperature optimum of 20 ℃,a pH optimum of 2.4,and Km value of 1.31 mM towards ABTS at pH 2.4 and 30 ℃.The ranking of its degradative activity towards various aromatic substrates was ATBS > guaiacol > catechol > hydroquinone > pyrogallol > N,N-dimethy1-1,4-phenylenediamine.Cu2+ can enhance the enzyme activity of 10.8-14.6 fold in the ion concentration range of 1.25-10 mM.IBL manifested antiproliferative activities towards HepG2 and L1210 cells with IC50 values of 2.4 μM and 3.2 μM,respectively,but is devoid of inhibitory activity towards HIV-1 reverse transcriotase.
Inonotus baumii Laccase Enzyme purification Characterization Antiproliferative activity
QING-JUN CHEN JIAN SUN HE-XIANG WANG GUO-QING ZHANG
Key Laboratory of Urban Agriculture(North)of Ministry of Agriculture, College of Biological Sciences State Key Laboratory of Agro-Biotechnology and MOA Key Laboratory of Soil Microbiology, College of B Key Laboratory of Urban Agriculture(North)of Ministry of Agriculture, College of Biological Sciences
国际会议
The 7th International Medicinal Mushroom Congerence(第七届国际药用菌大会(IMMC7)
北京
英文
269-280
2013-08-26（万方平台首次上网日期，不代表论文的发表时间）