Differential interactions of halophilic and non-halophilic proteases with nanoparticles
Tóm tắt
Increase in the industrial use of nanomaterials and nanoparticles (NPs) make their release into the environment inevitable. This may lead to environmental contamination and exposure of the biological/ microbial diversity. Nanoparticles have been reported to impregnate the cells and interact with cellular biomolecules especially proteins and DNA, leading to nanotoxicity in many cases. The present work targets to study nanoparticle-protein interactions in-vitro, especially to assess their effects on extracellularly secreted enzymes. The primary extracellular enzymes viz. hydrolases and proteases could be the first to come in contact with environmentally released nanoparticles. Two halophilic proteases from Geomicrobium sp. EMB2 and Bacillus sp. EMB9 and one non-halophilic protease, subtilisin from Bacillus licheniformis have been investigated for their interaction with silver and zinc oxide nanoparticles as model systems. The activities of Geomicrobium sp. and Bacillus sp. protease were unaffected while that of non-halophilic subtilisin was lost by 70% and 30% in presence of Ag and ZnO nanoparticles respectively. The secondary and tertiary structure of halophilic proteases was unchanged on exposure to ZnO and Ag nanoparticles. Non-halophilic protease showed significant loss in α-helical structure with changes in the microenvironment of the protein as observed by CD and fluorescence spectroscopy. The greater stability and structural integrity may be attributed to higher negative charges on the surfaces of halophilic proteins. Halophilic extracellular proteases were more stable and did not lose proteolytic activity. Their secondary structure remained unaffected by interaction with ZnO and Ag nanoparticles. Alterations in structure and loss of activity in non-halophilic protease have been quite prevalent on exposure to nanoparticles. The extracellular halophilic nanostable enzymes thus offer a promising robust system to counter nanotoxicity. A precise understanding of nanoparticle interaction with extracellular enzymes will pave the way for designing of novel enzymes and creating appropriate system to protect microbial diversity against nanoparticle disposal.
Tài liệu tham khảo
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