毛栓菌漆酶的分离纯化及酶学性质研究

对毛栓菌产漆酶的分离、纯化及酶学性质进行研究。粗酶液经硫酸铵盐析、透析、DEAE-Sepharose柱层析,得到2种漆酶同工酶LacA和LacB。LacA和LacB回收率分别为17.1%和2.74%。SDS-PAGE电泳测得2漆酶的分子量分别为54.6 ku和7.7 ku;LacA和LacB最适作用温度分别为50℃和60℃;最适反应pH值分别为4.5和4.0;Cu2＋、Mg2＋对LacA有激活作用,对LacB影响不大;Ag＋对LacA和LacB表现为完全抑制;Fe3＋对LacA和LacB有一定的抑制作用;Ca2＋、Mn2＋、K＋、Na＋、Zn2＋对LacA和LacB影响不大。DTT、EDTA、DMSO、SDS对酶均有不同程度的抑制作用,且随其浓度的升高抑制作用增强。

Isolation, Purification and Enzymatic Features of Laccase from Trametes trogii

Isolation,purification and the features of laccase from Trametes trogii were studied.Crude fermented broth was purified using salting-out with ammonium sulphate,dialysis,and DEAE-Sepharose column chromatography and obtained laccase isoenzyme LacA and LacB.The recovery of LacA and LacB were 17.1% and 2.74% respectively.The molecular weights of LacA and LacB were 54.6 ku and 17.7 ku respectively determined by SDS-PAGE electrophoresis.The optimal reaction temperature of LacA and LacB were 50 ℃ and 60 ℃ respectively,their optimal reaction pH were 4.5 and 4.0 respectively,Cu2+ and Mg2+ had activation on LacA,but had little effect on LacB,Ag+ completely inhibited both LacA and LacB,Fe3+ had a certain inhibition both on of LacA and LacB,Ca2+,Mn2+,K+,Na+,Zn2+ had not much effect both on LacA and LacB.DTT,EDTA,DMSO,SDS had different degree inhibition on activity of LacA and LacB,and the inhibition became stronger as the concentration raised.