Adsorption of peroxidase on Celite 545 directly from ammonium sulfate fractionated white radish (Raphanus sativus) proteins.
This paper demonstrates the direct immobilization of peroxidase from ammonium sulfate fractionated white radish proteins on an inorganic support, Celite 545. The adsorbed peroxidase was crosslinked by using glutaraldehyde. The activity yield for white radish peroxidase was adsorbed on Celite 545 was 70% and this activity was decreased and remained 60% of the initial activity after crosslinking by glutaraldehyde. The pH and temperature-optima for both soluble and immobilized peroxidase was at pH 5.5 and 40 degrees C. Immobilized peroxidase retained higher stability against heat and water-miscible organic solvents. In the presence of 5.0 mM mercuric chloride, immobilized white radish peroxidase retained 41% of its initial activity while the free enzyme lost 93% activity. Soluble enzyme lost 61% of its initial activity while immobilized peroxidase retained 86% of the original activity when exposed to 0.02 mM sodium azide for 1 h. The K(m) values were 0.056 and 0.07 mM for free and immobilized enzyme, respectively. Immobilized white radish peroxidase exhibited lower V(max) as compared to the soluble enzyme. Immobilized peroxidase preparation showed better storage stability as compared to its soluble counterpart.
Satar
Rukhsana
R
Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, UP, India.
Husain
Qayyum
Q
D000327:Adsorption; D000645:Ammonium Sulfate / Q000737:chemistry*; D001709:Biotechnology / Q000379:methods*; D003432:Cross-Linking Reagents / Q000737:chemistry; D007692:Diatomaceous Earth / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008627:Mercuric Chloride / Q000737:chemistry; D010544:Peroxidases / Q000737:chemistry*; D031224:Raphanus; D019810:Sodium Azide / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D013696:Temperature
D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D012997:Solvents; D008627:Mercuric Chloride; D007692:Diatomaceous Earth; D019810:Sodium Azide; D010544:Peroxidases; D000645:Ammonium Sulfate