Preparation and properties of RNase T2 immobilized on concanavalin A-sepharose.
Partially purified RNase T2 (EC 2.7.7.17) from Aspergillus oryzae was bound through its carbohydrate moiety to Concanavalin A-Sepharose. The retention of activity was high, ranging from 70% at low enzyme load to approximately 9% at high enzyme load. Though there was no change in the pH and temperature optima, the pH stability and the Km decreased after immobilization. Compared to the soluble enzyme, the immobilized RNase T2 showed enhanced temperature stability and more resistance to metal ions. Both soluble and immobilized enzymes were stable to 8 M urea. On repeated use, the bound enzyme retained more than 60% of its initial activity after six cycles.
Reddy
L G
LG
Division of Biochemical Sciences, National Chemical Laboratory, Pune, India.
Shankar
V
V
D001236:Aspergillus oryzae / Q000201:enzymology; D004722:Endoribonucleases / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D012685:Sepharose / Q000031:analogs & derivatives; D012995:Solubility; D013696:Temperature; D014508:Urea
D004800:Enzymes, Immobilized; D008670:Metals; C028326:concanavalin A-sepharose; D014508:Urea; D012685:Sepharose; D004722:Endoribonucleases; C022543:ribonuclease T(2)