Our Group organises 3000+ Global Conferenceseries Events every year across USA, Europe & Asia with support from 1000 more scientific Societies and Publishes 700+ Open Access Journals which contains over 50000 eminent personalities, reputed scientists as editorial board members.
Among lignocellulolytic enzymes, laccases (EC 1.10.3.2) are the most versatile, broadly specific and well-studied enzyme
with wide range of biotechnological potential. Putative laccase (BpCotA) from Bacillus pumilus MK001 was cloned
and expressed in E. coli BL21 (DE3) which displayed molecular weight of ~64 kDa, analyzed by SDS-PAGE and western
blotting. In addition to soluble bioactive fraction, inactive inclusion body (IB) fraction was also harvested and refolded under
optimized conditions resulting in 65% refolding efficiency. As predicted by far UV thermal CD spectra, increase in �?²-sheets of
the protein structure was recorded after thermal induction, investigated as one of the potent reason for higher thermostability
of the protein. Multiple sequence alignment with other similar proteins revealed the occurrence of highly conserved catalytic
residues. The 3D structure of BpCotA was constructed through homology modeling (Modeler 9.11). The structure was refined
and validated for stereochemical quality by using PROCHECK, ERRAT, Verify 3D and PROSA servers. The best selected model
with minimized energy was examined for superimposition with the crystal structure of CotA from B. subtilis which showed
0.33 RMSD value. The potential binding affinities of BpCotA was observed with ferulic acid, vanillic acid, vanillin and caffeic
acid involving flexible docking studies, AUTODOCK 4.2 and Hex showed that both ferulic acid and vanillin docked well in
the vicinity of predictive active site of recombinant protein. Further, in vitro action of enzyme was analyzed with respective
substrates and predicted interactions were confirmed by HPLC generated degradation spectra. Elucidation of thermostability,
interactions with phenolics and refolding of IB fraction will deduce the biotechnological applicability of BpCotA.