Multicopper oxidase (MCO) laccase from Stropharia sp. ITCC-8422: an apparent authentication using integrated experimenta
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ORIGINAL ARTICLE
Multicopper oxidase (MCO) laccase from Stropharia sp. ITCC‑8422: an apparent authentication using integrated experimental and in silico analysis Komal Agrawal1 · Jata Shankar2 · Pradeep Verma1 Received: 27 May 2020 / Accepted: 17 August 2020 © King Abdulaziz City for Science and Technology 2020
Abstract In the present study, specificity of laccase from Stropharia sp. ITCC-8422 against various substrates, i.e. 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,6-dimethoxyphenol (DMP), guaiacol (GCL) and syringaldazine (SYZ) was determined. It exhibited maximum affinity against ABTS, followed by DMP and negligible activity for GCL and SYZ. As the concentration of substrate increased from 0.5 to 1.5 mM (ABTS) and 1 to 5 mM (DMP), the activity increased from 301.1 to 567.8 U/L and 254.4 to 436.2 U/L. Further, quadrupole time-of-flight liquid chromatography mass spectrometry (QTOF-LCMS) analysis of the extracellular proteome of Stropharia sp. ITCC-8422 identified eighty-four (84) extracellular proteins. The peptide sequence for the enzyme of interest exhibited sequence similarity with laccase-5 of Trametes pubescens. Using high molecular mass sequence of laccase-5, the protein structure of laccase was modelled and binding energy of laccase with four substrates, i.e. ABTS (− 5.65), DMP (− 4.65), GCL (− 4.66) and SYZ (− 5.5) was determined using autodock tool. The experimental and in silico analyses revealed maximum activity of laccase and lowest binding energy with ABTS. Besides, laccase was purified and it exhibited 2.1-fold purification with purification yield of 20.4% and had stability of 70% at pH 5–9 and 30–40 ℃. In addition, the bioremediation potential of laccase was explored by in silico analysis, where the binding energy of laccase with alizarin cyanine green was − 6.37 and both in silico work and experimental work were in agreement. Keywords Laccase · Docking · In silico · Alizarin cyanine green · Bioremediation
Introduction The multicopper oxidases (MCOs) consist of a family of enzymes which couples the reduction of molecular oxygen to water. The MCO superfamily consists of laccase (EC 1.10.3.2), ascorbate oxidase (EC 1.10.3.3), ferroxidase (EC Electronic supplementary material The online version of this article (https://doi.org/10.1007/s13205-020-02399-8) contains supplementary material, which is available to authorized users. * Pradeep Verma [email protected]; [email protected] 1
Bioprocess and Bioenergy Laboratory, Department of Microbiology, Central University of Rajasthan, NH‑8, Bandarsindari, Kishangarh, Ajmer 305817, India
Genomics Laboratory, Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Solan 173234, Himachal Pradesh, India
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1.16.3.1), nitrite reductase (EC 1.7.2.1), bilirubin oxidase (EC 1.3.3.5) and ceruloplasmin (EC 1.16.3.1) where laccase has broad substrate specificity, e.g. polyamines and certain inorganic compounds (Baldrian 2005; Madhavi and Lele 2009; Janusz et al. 2020). Laccase is a
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