重组血红密孔菌漆酶的纯化、酶学性质及染料脱色研究

为了研究血红密孔菌漆酶重组表达后的酶学性质和染料脱色效果，利用超滤、阴离子交换层析和凝胶层析从毕赤酵母发酵液中纯化出重组漆酶。结果表明:重组漆酶分子质量为62.8 kD，氧化底物2,2′--连氮--二（3--乙基苯并噻唑--6--磺酸）（ABTS）、丁香醛连氮和2,6--二甲氧基苯酚的最适pH分别为2.2、4.2和4.0，动力学常数Km分别为20.73、48.48和1 252.39 μmol/L；该漆酶在有机溶剂中的活性稳定性:甲醇乙醇丙酮二甲基亚砜；在铜离子和铝离子中酶活比较稳定，汞离子对酶活有较强的抑制作用；纯化的重组漆酶在无介体时可有效脱色RB亮蓝，但对靛红和结晶紫的脱色较为缓慢；当添加介体ABTS或紫脲酸时，可在短时间内实现对靛红和结晶紫的脱色，显示出该重组漆酶在染料废水处理中具有较好的应用前景。 

Purification,characterization and dye decolorization of a recombinant Pycnoporus sanguineus laccase

To study the characterization and dye decolorization ability of a recombinant Pycnoporus sanguineus laccase,the enzyme was purified from the fermentation liquid of Pichia pastoris using ultrafiltration,anion-exchange chromatography and gel filtration.The results showed that the recombinant laccase had a molecular weight of around 62.8 kD.It oxidized the substrates 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS),syringaldazine and 2,6-dimethoxyphenol at optimum pH of 2.2,4.2 and 4.0,with Km valu...