caa a22 4500 605507678 CHVBK 20210128100633.0 cr unu---uuuuu 210128e20150701xx s 000 0 eng 10.1007/s00775-015-1266-2 doi (NATIONALLICENCE)springer-10.1007/s00775-015-1266-2 Participatory role of zinc in structural and functional characterization of bioremediase: a unique thermostable microbial silica leaching protein [Elektronische Daten] [Trinath Chowdhury, Manas Sarkar, Biswadeep Chaudhuri, Brajadulal Chattopadhyay, Umesh Halder] A unique protein, bioremediase (UniProt Knowledgebase Accession No.: P86277), isolated from a hot spring bacterium BKH1 (GenBank Accession No.: FJ177512), has shown to exhibit silica leaching activity when incorporated to prepare bio-concrete material. Matrix-assisted laser desorption ionization mass spectrometry analysis suggests that bioremediase is 78% homologous to bovine carbonic anhydrase II though it does not exhibit carbonic anhydrase-like activity. Bioinformatics study is performed for understanding the various physical and chemical parameters of the protein which predicts the involvement of zinc encircled by three histidine residues (His94, His96 and His119) at the active site of the protein. Isothermal titration calorimetric-based thermodynamic study on diethyl pyrocarbonate-modified protein recognizes the presence of Zn2+ in the enzyme moiety. Exothermic to endothermic transition as observed during titration of the protein with Zn2+ discloses that there are at least two binding sites for zinc within the protein moiety. Addition of Zn2+ regains the activity of EDTA chelated bioremediase confirming the presence of extra binding site of Zn2+ in the protein moiety. Revival of folding pattern of completely unfolded urea-treated protein by Zn2+ explains the participatory role of zinc in structural stability of the protein. Restoration of the λ max in intrinsic fluorescence emission study of the urea-treated protein by Zn2+ similarly confirms the involvement of Zn in the refolding of the protein. The utility of bioremediase for silica nanoparticles preparation is observed by field emission scanning electron microscopy. Graphical abstract: SBIC, 2015 Bioinformatics nationallicence Calorimetry nationallicence Circular dichroism nationallicence MALDI-MS nationallicence Metalloprotein nationallicence Zinc nationallicence Chowdhury Trinath Department of Physics, Jadavpur University, 700032, Kolkata, India aut Sarkar Manas Department of Physics, Jadavpur University, 700032, Kolkata, India aut Chaudhuri Biswadeep Department of Physics, Jadavpur University, 700032, Kolkata, India aut Chattopadhyay Brajadulal Department of Physics, Jadavpur University, 700032, Kolkata, India aut Halder Umesh Department of Chemistry, Jadavpur University, 700032, Kolkata, India aut JBIC Journal of Biological Inorganic Chemistry Springer Berlin Heidelberg 20/5(2015-07-01), 791-803 0949-8257 20:5<791 2015 20 775 https://doi.org/10.1007/s00775-015-1266-2 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00775-015-1266-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Chowdhury Trinath Department of Physics, Jadavpur University, 700032, Kolkata, India aut NATIONALLICENCE 700 1- Sarkar Manas Department of Physics, Jadavpur University, 700032, Kolkata, India aut NATIONALLICENCE 700 1- Chaudhuri Biswadeep Department of Physics, Jadavpur University, 700032, Kolkata, India aut NATIONALLICENCE 700 1- Chattopadhyay Brajadulal Department of Physics, Jadavpur University, 700032, Kolkata, India aut NATIONALLICENCE 700 1- Halder Umesh Department of Chemistry, Jadavpur University, 700032, Kolkata, India aut NATIONALLICENCE 773 0- JBIC Journal of Biological Inorganic Chemistry Springer Berlin Heidelberg 20/5(2015-07-01), 791-803 0949-8257 20:5<791 2015 20 775