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Heterologous expression and characterization of laccase 2 from Coprinopsis cinerea capable of decolourizing different recalcitrant dyes

文献类型： 外文期刊

作者： Tian, Yong-Sheng 1 ; Xu, Hu 1 ; Peng, Ri-He 1 ; Yao, Quan-Hong 1 ; Wang, Rong-Tan 2 ;

作者机构： 1.Shanghai Acad Agr Sci, Inst Agrobiotechnol Res, Shanghai, Peoples R China

2.Shanghai Ruifeng Agr Sci & Technol Co Ltd, Shanghai, Peoples R China

关键词： laccase;basidiomycete;Coprinopsis cinerea Okayama-7;130;methyl orange;crystal violet;malachite green

期刊名称：BIOTECHNOLOGY & BIOTECHNOLOGICAL EQUIPMENT （ 影响因子：1.632； 五年影响因子：2.029 ）

ISSN：

年卷期：

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收录情况： SCI

摘要： The gene (CcLcc2) encoding laccase from the basidiomycete Coprinopsis cinerea Okayama-7 #130 was synthesized by polymerase chain reaction-based two-step DNA synthesis, and heterologously expressed in Pichia pastoris. The recombinant protein was purified by ammonium sulphate precipitation and nickel nitrilotriacetic acid chromatography. The molecular mass of CcLcc2 was estimated to be 54 kDa by denaturing polyacrylamide gel electrophoresis. The optimum pH and temperature for laccase catalysis for the oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonate) (ABTS) were 2.6 and 45 degrees C, respectively. The Km values of the enzyme towards the substrates ABTS, 2,6-dimethoxyphenol (2,6-DMP) and guaiacol were 0.93, 1.02 and 28.07 mmol.L-1, respectively. The decolourization of methyl orange, crystal violet and malachite green, commonly used in the textile industry, was assessed. The decolourization percentage of crystal violet and malachite green was 80% after 4 h of reaction, and that of methyl orange was 50% at 4 h. These results show that the CcLcc2 has enormous potential for the decolourization of highly stable triphenylmethane dyes.

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