Polymer-assisted iron oxide magnetic nanoparticle immobilized keratinase.
Nanotechnology holds the prospect for avant-garde changes to improve the performance of materials in various sectors. The domain of enzyme biotechnology is no exception. Immobilization of industrially important enzymes onto nanomaterials, with improved performance, would pave the way to myriad application-based commercialization. Keratinase produced by Bacillus subtilis was immobilized onto poly(ethylene glycol)-supported Fe3O4 superparamagnetic nanoparticles. The optimization process showed that the highest enzyme activity was noted when immobilized onto cyanamide-activated PEG-assisted MNP prepared under conditions of 25 degrees C and pH 7.2 of the reaction mixture before addition of H2O2 (3% w/w), 2% (w/v) PEG(6000) and 0.062:1 molar ratio of PEG to FeCl2 x 4H2O. Further statistical optimization using response surface methodology yielded an R2 value that could explain more than 94% of the sample variations. Along with the magnetization studies, the immobilization of the enzyme onto the PEG-assisted MNP was characterized by UV, XRD, FTIR and TEM. The immobilization process had resulted in an almost fourfold increase in the enzyme activity over the free enzyme. Furthermore, the immobilized enzyme exhibited a significant thermostability, storage stability and recyclability. The leather-industry-oriented application of the immobilized enzyme was tested for the dehairing of goat-skin.
Advanced Polymer and Nanomaterial Laboratory, Department of Chemical Sciences, Tezpur University, Tezpur-784028, Assam, India.
Rocktotpal
R
Konwarh
Niranjan
N
Karak
Sudhir Kumar
SK
Rai
Ashis Kumar
AK
Mukherjee
D000818:Animals; D001412:Bacillus subtilis / Q000201:enzymology; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D004574:Electromagnetic Fields; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D017743:Equipment Reuse; D005290:Ferric Compounds / Q000737:chemistry*; D006041:Goats; D006197:Hair / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000737:chemistry*; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry*; D012044:Regression Analysis; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D005290:Ferric Compounds; C000499:ferric oxide; D011092:Polyethylene Glycols; D006861:Hydrogen Peroxide; D010447:Peptide Hydrolases; C035855:keratinase