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Biochemical Characteristics of Three Laccase Isoforms from the Basidiomycete Pleurotus nebrodensis

文献类型： 外文期刊

作者： Yuan, Xianghe 1 ; Tian, Guoting 3 ; Zhao, Yongchang 3 ; Zhao, Liyan 4 ; Wang, Hexiang 1 ; Ng, Tzi Bun 5 ;

作者机构： 1.China Agr Univ, State Key Lab Agrobiotechnol, Beijing 100193, Peoples R China

2.China Agr Univ, Dept Microbiol, Beijing 100193, Peoples R China

3.Yunnan Acad Agr Sci, Inst Biotechnol & Germplasm Resource, Kunming 650223, Peoples R China

4.Nanjing Agr Univ, Coll Food Sci & Technol, Nanjing 210095, Jiangsu, Peoples R China

5.Chinese Univ Hong Kong, Sch Biomed Sci, Fac Med, Shatin, Hong Kong, Peoples R China

关键词： laccase isoenzymes;enzyme characterization;biochemical properties comparison;Pleurotus nebrodensis

期刊名称：MOLECULES （ 影响因子：4.411； 五年影响因子：4.587 ）

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收录情况： SCI

摘要： The characterization of three laccase isoforms from Pleurotus nebrodensis is described.Isoenzymes Lac1, Lac2 and Lac3 were purified to homogeneity using ion exchange chromatographyon DEAE-cellulose, CM-cellulose and Q-Sepharose and a gel filtration step on Superdex 75.The molecular weights of the purified laccases were estimated to be 68, 64 and 51 kDa, respectively.The isoenzymes demonstrated the same optimum pH at 3.0 but slightly different temperature optima:50–60 C for Lac1 and Lac3 and 60 C for Lac2. Lac2 was always more stable than the other two isoforms and exposure to 50 C for 120 min caused 30% loss in activity. Lac2 was relatively less stable than the other two isoforms when exposed to the pH range of 3.0–8.0 for 24 h, but inactivation only occurred initially, with around 70% residual activity being maintained during the whole process.Oxidative ability towards aromatic compounds varied substantially among the isoforms and each of them displayed preference toward some substrates. Kinetic constants (Km, Kcat) were determined by using a 2,21-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assay,with Lac3 showing the best affinity and Lac2 displaying the highest catalytic efficiency. Amino acid sequences from peptides derived from digestion of isoenzymes showed great consistency with laccases in the databases.

  • 相关文献

[1]Degradation of dyes using crude extract and a thermostable and pH-stable laccase isolated from Pleurotus nebrodensis. Yuan, Xianghe,Wang, Hexiang,Yuan, Xianghe,Wang, Hexiang,Tian, Guoting,Zhao, Yongchang,Zhao, Liyan,Ng, Tzi Bun. 2016

[2]Purification and characterization of a novel laccase from the mushroom pleurotus nebrodensis. Wang, He-Xiang,Wang, He-Xiang,Ng, Tzi Bun,Zhang, Guo-Qing,Tian, Guo-Ting.

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