Abstract

The oxidation activity of multicopper-oxidases overlaps with different substrates of laccases and bilirubin oxidases, thus in the present study an integrated approach of bioinformatics using homology modeling, docking, and experimental validation was used to confirm the type of multicopper-oxidase in Myrothecium verrucaria ITCC-8447. The result of peptide sequence of M. verrucaria ITCC-8447 enabled to predict the 3 D-structure of multicopper-oxidase. It was overlapped with the structure of laccase and root mean square deviation (RMSD) was 1.53 Å for 533 and, 171 residues. The low binding energy with azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) (-5.64) as compared to bilirubin (-4.39) suggested that M. verrucaria ITCC-8447 have laccase-like activity. The experimental analysis confirmed high activity with laccase specific substrates, phenol (18.3 U/L), ampyrone (172.4 U/L) and, ampyrone phenol coupling (50 U/L) as compared to bilirubin oxidase substrate bilirubin (16.6 U/L). In addition, lowest binding energy with ABTS (-5.64), syringaldazine SYZ (-4.83), guaiacol GCL (-4.42), and 2,6-dimethoxyphenol DMP (-4.41) confirmed the presence of laccase. Further, complete remediation of two hazardous model pollutants i.e., phenol and resorcinol (1.5 mM) after 12 h of incubation and low binding energy of −4.32 and, −4.85 respectively confirmed its removal by laccase. The results confirmed the presence of laccase in M. verrucaria ITCC-8447 and its effective bioremediation potential.

摘要

多铜氧化酶的氧化活性与漆酶和胆红素氧化酶的不同底物重叠，因此，在本研究中，采用同源建模，对接和生物验证的一种综合生物信息学方法，以证实疣粒黑夜蛾ITCC中的多铜氧化酶的类型。-8447。疣状支原体ITCC-8447的肽序列结果使得能够预测多铜氧化酶的3D结构。它与漆酶的结构重叠，533个残基和171个残基的均方根偏差（RMSD）为1.53Å。与相比，胆红素连氮基-双（3-乙基苯并噻唑-6-磺酸）（ABTS）（-5.64）的低结合能（-4.39）建议，M.疣ITCC-8447具有漆酶样活性。实验分析证实，与胆红素氧化酶底物胆红素（16.6 U / L）相比，漆酶特异性底物，苯酚（18.3 U / L），氨苯醌（172.4 U / L）和氨苯醌苯酚偶联（50 U / L）具有较高的活性。 。此外，与ABTS（-5.64），丁香嗪SYZ（-4.83），愈创木酚GCL（-4.42）和2,6-二甲氧基苯酚DMP（-4.41）的最低结合能证实了漆酶的存在。此外，在孵育12小时后，对两种有害模型污染物，即苯酚和间苯二酚（1.5 mM）的完全修复，以及-4.32和-4.85的低结合能，分别证实了其被漆酶去除。结果证实漆酶存在于疣状支原体ITCC-8447中，并具有有效的生物修复潜力。