title: Expression and characteristics of manganese peroxidase from Ganoderma lucidum in Pichia pastoris and its application in the degradation of four dyes and phenol.
abstract: Manganese peroxidase (MnP) of white rot basidiomycetes, an extracellular heme enzyme, is part of a peroxidase superfamily that is capable of degrading the different phenolic compounds. Ganoderma, a white rot basidiomycete widely distributed worldwide, could secrete lignin-modifying enzymes (LME), including laccase (Lac), lignin peroxidases (LiP) and MnP.
  
  
  
  After the selection of a G. lucidum strain from five Ganoderma strains, the 1092 bp full-length cDNA of the MnP gene, designated as G. lucidum MnP (GluMnP1), was cloned from the selected strain. We subsequently constructed an eukaryotic expression vector, pAO815:: GlMnP, and transferred it into Pichia pastoris SMD116. Recombinant GluMnP1 (rGluMnP1) was with a yield of 126 mg/L and a molecular weight of approximately 37.72 kDa and a specific enzyme activity of 524.61 U/L. The rGluMnP1 could be capable of the decolorization of four types of dyes and the degradation of phenol. Phenol and its principal degradation products including hydroquinone, pyrocatechol, resorcinol, benzoquinone, were detected successfully in the experiments.
  
  
  
  The rGluMnP1 could be effectively expressed in Pichia pastoris and with a higher oxidation activity. We infer that, in the initial stages of the reaction, the catechol-mediated cycle should be the principal route of enzymatic degradation of phenol and its oxidation products. This study highlights the potential industrial applications associated with the production of MnP by genetic engineering methods, and the application of industrial wastewater treatment.
authors.affiliation: Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, and Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education, and School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, People's Republic of China.
authors.forename: Hui
authors.initials: H
authors.lastname: Xu
authors.affiliation: Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, and Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education, and School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, People's Republic of China.
authors.forename: Meng-Yuan
authors.initials: MY
authors.lastname: Guo
authors.affiliation: Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, and Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education, and School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, People's Republic of China.
authors.forename: Yan-Hua
authors.initials: YH
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, People's Republic of China. xhbai@sjtu.edu.cn.
authors.forename: Xiao-Hui
authors.initials: XH
authors.lastname: Bai
authors.affiliation: Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, and Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education, and School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, People's Republic of China. xuanweizhou@sjtu.edu.cn.
authors.forename: Xuan-Wei
authors.initials: XW
authors.lastname: Zhou
mesh_terms: D001673:Biodegradation, Environmental; D003001:Cloning, Molecular / Q000379:methods; D004396:Coloring Agents / Q000737:chemistry* / Q000302:isolation & purification; D004789:Enzyme Activation; D010544:Peroxidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019800:Phenol / Q000737:chemistry* / Q000302:isolation & purification; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020940:Reishi / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D013045:Species Specificity; D014874:Water Pollutants, Chemical / Q000737:chemistry* / Q000302:isolation & purification; D018508:Water Purification / Q000379:methods
chemical_list: D004396:Coloring Agents; D011994:Recombinant Proteins; D014874:Water Pollutants, Chemical; D019800:Phenol; D010544:Peroxidases; C051129:manganese peroxidase
keywords: Degradation; Ganoderma lucidum; Manganese peroxidase; Phenolic compound; Yeast expression system