Purification and characterization of two thermostable laccases from Pycnoporus sanguineus and potential role in degradation of endocrine disrupting chemicals

• DOI: 10.1016/j.molcatb.2014.06.006
• Publication Date: 2014-07-02T10:50:30Z
• Authors: Leticia I. Ramíre...; Charles Junghanns; Nancy Ornelas-Soto; Diana L. Cárdenas...; Carlos Hernández-...; Philippe Demarche; Estelle Enaud; Raúl García-Morales; Spiros N. Agathos; Roberto Parra
• Keywords: Thermostability; ABTS; Guaiacol

In this study, the purification and characterization of two thermostable laccases produced by Pycnoporus sanguineus CS43 (LacI LacII) were performed. Also, their biotechnological potential was assessed through degradation endocrine disrupting chemicals (EDCs). Laccases purified ultrafiltration, ion exchange (IEX) hydrophobic interaction chromatography achieving specific activities close to 285 U mg−1. The molecular weights LacI LacII, determined SDS-electrophoresis, 68 66 kDa, respectively. Both showed high amino acid sequence similarity (91%) between them thermostability, at 50 60 °C (half-lives 277.7 18 h for LacI, 35.8 2.25 LacII). isoforms oxidized common laccase substrates such as 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonate (ABTS), 2,6-dimethoxyphenol (DMP) guaiacol acidic pH conditions. ABTS most efficient substrate, showing specificity constants 74,816 36,746 mM−1 s−1 Lac I Michaelis (Km) 6.9 12.2 μM respectively 3. remained active concentrations organic solvents (acetonitrile, ethanol acetone), with an IC50 (v/v) >64%, 55% 47% 33%, 52% 31% LacII tested degrade EDCs, nonylphenol triclosan, more than 95% removal after 8 treatment 100 U/L 5 means HPLC. unique Michaelis–Menten kinetic parameters, solvent tolerance demonstrated P. render promising candidates industrial applications. exerted a higher thermal stability, against inhibitors catalyst DMP LacII.