Purification and biochemical characterization of two laccase isoenzymes isolated from Trichoderma harzianum S7113 and its application for bisphenol A degradation

Microbial Cell Factories

Research

Abdelmageed M. Othman¹  Mohamed Abdelraof¹  Alshaimaa M. Elsayed²  Ghada S. A. Abdel Karim²  Mohamed Mahmoud³ 

[1] Microbial Chemistry Department, Biotechnology Research Institute, National Research Centre, Dokki, 12622, Giza, Egypt;Molecular Biology Department, Biotechnology Research Institute, National Research Centre, Dokki, 12622, Giza, Egypt;Water Pollution Research Department, National Research Centre, 33 El-Buhouth St., Dokki, 12622, Giza, Egypt;

关键词: Laccase;    Purification;    Biochemical characterization;    Isoenzymes;    Bisphenol A;    Degradation;   

DOI  :  10.1186/s12934-022-02011-z

 received in 2022-12-03, accepted in 2022-12-23,  发布年份 2022

来源: Springer

【 摘 要 】

Two laccase isoenzymes (LacA and LacB) were isolated from a novel Trichoderma harzianum S7113 isolate employing ammonium sulfate precipitation, Sephadex G100, and DEAE Sepharose ion exchange chromatography. The molecular weights of the purified LacA and LacB laccases were estimated to be 63 and 48 kDa, respectively. The two isoenzymes had their optimum activities at the same temperature (50 °C), but at slightly different pH values (pH 3.0 for LacA and pH 2.5 for LacB). LacA and LacB had the same thermal stability at 40 °C and pH stability at pH 9.0. The two isoenzymes also showed a high level of specific activity toward ABTS, where the Km values of LacA and LacB were 0.100 and 0.065 mM, whereas their Vmax values were 0.603 and 0.182 µmol min−1, respectively. LacA and LacB catalytic activity was stimulated by Mg2+, Zn2+, K+, and Ni2+, whereas it was inhibited by Hg2+ and Pb2+, β-mercaptoethanol, EDTA, and SDS, and completely inhibited by sodium azide. Our findings indicate that purified laccase has a promising capacity for bisphenol A (BPA) bioremediation across a broad pH range. This finding opens up new opportunities for the commercialization of this technique in a variety of biotechnology-based applications, particularly for removing endocrine chemicals from the environment.

【 授权许可】

CC BY   
© The Author(s) 2023

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