title: Purification and characterization of a laccase from Coprinopsis cinerea in Pichia pastoris.
abstract: A modified laccase gene, CcLCC6, from Coprinopsis cinerea was chemically synthesized according to the yeast codon bias and expressed in Pichia pastoris. The main properties of laccase, effects of ions and inhibitors, and optimal condition for decolouring malachite green (MG) were investigated in this study. The optimal pH level and temperature of laccase are 3.0 and 40 °C, respectively. The metal ions Mn²⁺, Zn²⁺, Fe³⁺ and Al³⁺ could inhibit laccase activity, as well as 1 mM of sodium dodecyl sulphate and sodium thiosulphate. 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), as a mediator, was necessary in decolorizing MG. The optimal pH and temperature for MG decolorization were 3.0 and 50 °C, respectively. Approximately 0.02 μM recombinant laccase could effectively decolour 0.05 mM of MG in 1 h. CcLCC6I could inhibit the toxicity of MG to P. pastoris. This is the first report on the successful expression in P. pastoris of CcLCC6I and its enzymatic property. Laccase can also be considered as a candidate for treating industrial effluent containing MG.
authors.affiliation: Agro-Biotechnology Research Center, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China, wangbo_2009@sogou.com.
authors.forename: Bo
authors.initials: B
authors.lastname: Wang
authors.forename: Lijuan
authors.initials: L
authors.lastname: Wang
authors.forename: Yaqiu
authors.initials: Y
authors.lastname: Lin
authors.forename: Qing
authors.initials: Q
authors.lastname: Han
authors.forename: Jing
authors.initials: J
authors.lastname: Han
authors.forename: Jianjie
authors.initials: J
authors.lastname: Gao
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Tian
authors.forename: Wei
authors.initials: W
authors.lastname: Zhao
authors.forename: Rihe
authors.initials: R
authors.lastname: Peng
authors.forename: Quanhong
authors.initials: Q
authors.lastname: Yao
mesh_terms: D000363:Agaricales / Q000201:enzymology*; D001711:Biotransformation; D003001:Cloning, Molecular; D003116:Color; D004791:Enzyme Inhibitors / Q000032:analysis*; D004795:Enzyme Stability; D015870:Gene Expression; D005822:Genetic Vectors; D006863:Hydrogen-Ion Concentration; D007477:Ions / Q000378:metabolism*; D042845:Laccase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012394:Rosaniline Dyes / Q000378:metabolism*; D013696:Temperature
chemical_list: D004791:Enzyme Inhibitors; D007477:Ions; D011994:Recombinant Proteins; D012394:Rosaniline Dyes; C005095:malachite green; D042845:Laccase