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菌物学报  2015 

白灵侧耳漆酶分离纯化及其酶学性质研究

DOI: DOI:10.13346/j.mycosystema.140057, PP. 456-464

Keywords: 白灵侧耳,漆酶,分离纯化,酶活

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Abstract:

为了探索白灵侧耳Pleurotuseryngiivar.tuoliensis漆酶性质,以白灵侧耳菌株00485为试验材料,从发酵液中分离纯化得到胞外漆酶并对其酶学性质进行测定。纯化流程依次为DEAE-Cellulose阴离子交换层析,CM-Cellulose阳离子交换层析,SP-Sepharose强阳离子交换层析以及Superdex75凝胶过滤层析,获得胞外白灵侧耳漆酶(PnLac)。SDS-PAGE检测结果表明PnLac为65kDa的单一蛋白。PnLac经过胰蛋白酶水解得到3种肽段,经过NBCI-BLAST后发现它们与糙皮侧耳、环柄韧伞、刺芹侧耳等的漆酶具有同源性。底物为2,2?-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)时该种漆酶的最适反应温度和pH分别为50℃和3.0,Ca2+和Hg2+能够抑制它的活性,相反地Cu2+和Mn2+能够提高它的活性,米氏常数Km和Vmax分别是0.17mmol/L和1.76OD/min/U。

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