title: Novel magnetic cross-linked enzyme aggregates (magnetic CLEAs) of alpha amylase.
abstract: Novel magnetic cross-linked enzyme aggregates of alpha amylase were prepared by chemical cross-linking of enzyme aggregates with amino functionalized magnetite nanoparticles which can be separated from reaction mixture using magnetic field. Of the initially applied alpha amylase activity 100% was recovered in magnetic CLEAs, whereas only 45% was recovered in CLEAs due to the low content of Lys residues in alpha amylase. Scanning electron microscopy analysis showed that CLEAs and magnetic CLEAs were spherical structures. The CLEAs and magnetic CLEAs displayed a shift in optimal pH towards the acidic side, whereas optimal temperature of magnetic CLEAs was improved compared to free enzyme and CLEAs. Although V(max) of enzyme in CLEAs and magnetic CLEAs did not change, substrate affinity of the enzyme increased. The magnetic CLEAs also enhanced the thermal stability and storage stability. Moreover, the magnetic CLEAs retained 100% initial activity even after 6 cycles of reuse.
authors.affiliation: Department of Biotechnology Engineering, Kolhapur Institute of Technology's College of Engineering, Kolhapur 416 234, India. sachintalekar7@gmail.com
authors.forename: Sachin
authors.initials: S
authors.lastname: Talekar
authors.forename: Vishal
authors.initials: V
authors.lastname: Ghodake
authors.forename: Trupti
authors.initials: T
authors.lastname: Ghotage
authors.forename: Pavan
authors.initials: P
authors.lastname: Rathod
authors.forename: Priyant
authors.initials: P
authors.lastname: Deshmukh
authors.forename: Shamraja
authors.initials: S
authors.lastname: Nadar
authors.forename: Mosin
authors.initials: M
authors.lastname: Mulla
authors.forename: Mayur
authors.initials: M
authors.lastname: Ladole
mesh_terms: D001407:Bacillus / Q000201:enzymology; D011232:Chemical Precipitation; D003432:Cross-Linking Reagents / Q000378:metabolism*; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008280:Magnetics / Q000379:methods*; D020836:Protein Structure, Quaternary; D059027:Recycling; D013696:Temperature; D000516:alpha-Amylases / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D003432:Cross-Linking Reagents; D000516:alpha-Amylases