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Stability and Kinetic Properties of Immobilized Rhodotorula gracilis d ‐Amino Acid Oxidase
Author(s) -
Pilone Mirella S.,
Pollegioni Loredano,
Butò Simona
Publication year - 1992
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1992.tb00001.x
Subject(s) - d amino acid oxidase , chemistry , substrate (aquarium) , immobilized enzyme , oxidase test , enzyme , cofactor , chromatography , amino acid , alanine , amine gas treating , biochemistry , rhodotorula , stereochemistry , organic chemistry , yeast , biology , ecology
d ‐Amino acid oxidase form Rhodotorula gracilis has been immobilized with the use of amine reactive coupling chemistries. Coupling to Affi‐Gel 10 was very effective: yields of 50 to 60% were attained under optimal conditions with a specific activity of immobilized enzyme comparable to that of the free suspended preparation. No loss of the coenzyme FAD was observed under any experimental conditions. Immobilized d ‐amino acid oxidase exhibited a marked enhancement of stability against inactivation on heating, on pH shifts, and on denaturating agents; a relevant operational and storage stability was furthermore shown by the coupled system. Substrate specificity of the Affi‐Gel‐coupled enzyme was broader than for free enzyme; lowered apparent K m s and unchanged specific activity values for d ‐alanine, d ‐leucine, d ‐proline, d ‐valine, and cephalosporin C were determined for immobilized enzyme compared to the soluble form.