Purification, immobilization, and characterization of nattokinase on PHB nanoparticles.
In this study, nattokinase was purified from Bacillus subtilis using ion exchange chromatography and immobilized upon polyhydroxybutyrate (PHB) nanoparticles. A novel strain isolated from industrial dairy waste was found to synthesize polyhydroxyalkanoates (PHA) and the strain was identified as Brevibacterium casei SRKP2. PHA granules were extracted from 48 h culture and the FT-IR analysis characterized them as PHB, a natural biopolymer from B. casei. Nanoprecipitation by solvent displacement technique was used to synthesize PHB nanoparticles. PHB nanoparticles were characterized using transmission electron microscopy and particle size ranged from 100-125 nm. Immobilization of nattokinase upon PHB nanoparticles resulted in a 20% increase in the enzyme activity. Immobilization also contributed to the enhanced stability of the enzyme. Moreover, the activity was completely retained on storage at 4 degrees C for 25 days. The method has proven to be highly simple and can be implemented to other enzymes also.
Deepak
Venkataraman
V
Department of Biotechnology, Division of Molecular and Cellular Biology, Kalasalingam University, Anand Nagar, Krishnankoil 626190, Tamil Nadu, India.
Pandian
Suresh babu Ram Kumar
Sb
Kalishwaralal
Kalimuthu
K
Gurunathan
Sangiliyandi
S
D001412:Bacillus subtilis / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D053758:Nanoparticles / Q000737:chemistry*; D011091:Polyesters / Q000737:chemistry*; D054813:Polyhydroxyalkanoates / Q000737:chemistry*; D013381:Subtilisins / Q000302:isolation & purification*; D013696:Temperature
D004800:Enzymes, Immobilized; D011091:Polyesters; D054813:Polyhydroxyalkanoates; D013381:Subtilisins; C053771:nattokinase