title: Heterologous expression and characterization of laccase 2 fromCoprinopsis cinereacapable of decolourizing different recalcitrant dyes.
abstract: The gene (CcLcc2) encoding laccase from the basidiomyceteCoprinopsis cinereaOkayama-7 #130 was synthesized by polymerase chain reaction-based two-step DNA synthesis, and heterologously expressed inPichia 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 °C, respectively. TheKmvalues 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.
authors.affiliation: Institute of Agro-Biotechnology Research, Shanghai Academy of Agricultural Sciences , Shanghai , P.R.China.
authors.forename: Yong-Sheng
authors.initials: YS
authors.lastname: Tian
authors.affiliation: Institute of Agro-Biotechnology Research, Shanghai Academy of Agricultural Sciences , Shanghai , P.R.China.
authors.forename: Hu
authors.initials: H
authors.lastname: Xu
authors.affiliation: Institute of Agro-Biotechnology Research, Shanghai Academy of Agricultural Sciences , Shanghai , P.R.China.
authors.forename: Ri-He
authors.initials: RH
authors.lastname: Peng
authors.affiliation: Institute of Agro-Biotechnology Research, Shanghai Academy of Agricultural Sciences , Shanghai , P.R.China.
authors.forename: Quan-Hong
authors.initials: QH
authors.lastname: Yao
authors.affiliation: Shanghai Ruifeng Agricultural Science and Technology Co. Ltd , Shanghai , P.R.China.
authors.forename: Rong-Tan
authors.initials: RT
authors.lastname: Wang
keywords: Coprinopsis cinerea Okayama-7 #130; basidiomycete; crystal violet; laccase; malachite green; methyl orange