Purification and characterization of a novel laccase from Coprinus cinereus and decolorization of different chemically dyes.
Laccase is a blue copper oxidase with multiple copper ions and widely distributed in higher plant and fungi. To date, numerous fungal laccases have been reported by many researchers. In present work, a new laccase gene, named CcLCC5I, from Coprinus cinereus was synthesized chemically according to the yeast bias codon and integrated into Pichia pastoris GS115 genome by electroporation. SDS-PAGE analysis showed that the recombinant laccase has a molecular mass of approximately 56.8 kDa. Its biochemical properties was carried out using substrate 2-2(')-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS). It was showed that the optimum pH and temperature of the laccase is 3.0 and 55 °C, respectively. Except for copper ions, most metal ions inhibited the laccase activity at a high concentration about 10 mM. Sodium sulfite can also highly inhibit laccase activity whereas EDTA had no inhibitory effect on the laccase activity. The CcLCC5I have high ability to decolor not only azo but also aryl methane dyes. The recombinant laccase decolored 44.6 % orange G, 54.8 % Crystal Violet, and 87.2 % Malachite green at about 2.6 h. The novel laccase may be a good candidate for breeding engineering strains used in the treatment of industrial effluent containing azo and aryl methane dyes.
College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China.
Yaqiu
Y
Lin
Zhen
Z
Zhang
Yongsheng
Y
Tian
Wei
W
Zhao
Bo
B
Zhu
Zhisheng
Z
Xu
Rihe
R
Peng
Quanhong
Q
Yao
D001391:Azo Compounds / Q000737:chemistry; D052160:Benzothiazoles / Q000737:chemistry; D002614:Chelating Agents / Q000737:chemistry; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004396:Coloring Agents / Q000737:chemistry*; D003300:Copper / Q000737:chemistry; D003302:Coprinus / Q000201:enzymology*; D004492:Edetic Acid / Q000737:chemistry; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D005813:Genes, Synthetic; D005840:Gentian Violet / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D042845:Laccase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D010843:Pichia; D011994:Recombinant Proteins; D012394:Rosaniline Dyes / Q000737:chemistry; D013447:Sulfites / Q000737:chemistry; D013451:Sulfonic Acids / Q000737:chemistry; D013696:Temperature; D018508:Water Purification
D001391:Azo Compounds; D052160:Benzothiazoles; D002614:Chelating Agents; D004396:Coloring Agents; D005656:Fungal Proteins; D011994:Recombinant Proteins; D012394:Rosaniline Dyes; D013447:Sulfites; D013451:Sulfonic Acids; C005095:malachite green; C008710:Orange G; C002502:2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid; D003300:Copper; D004492:Edetic Acid; D042845:Laccase; D005840:Gentian Violet; C025026:sodium sulfite