Simultaneous purification and reversible immobilization of D-amino acid oxidase from Trigonopsis variabilis on a hydrophobic support

Citation
Sf. D'Souza et A. Deshpande, Simultaneous purification and reversible immobilization of D-amino acid oxidase from Trigonopsis variabilis on a hydrophobic support, APPL BIOC B, 95(2), 2001, pp. 83-92
Citations number
33
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biotecnology & Applied Microbiology","Biochemistry & Biophysics
Journal title
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
ISSN journal
0273-2289 → ACNP
Volume
95
Issue
2
Year of publication
2001
Pages
83 - 92
Database
ISI
SICI code
0273-2289(200108)95:2<83:SPARIO>2.0.ZU;2-8
Abstract
Purification and reversible immobilization Of D-amino acid oxidase from Tri gonopsis variabilis could be simultaneously accomplished by hydrophobic int eraction on Phenyl Sepharose CL-4B in the presence of 50 mM pyrophosphate b uffer (pH 8.5). The presence of a high salt concentration of 2 M, which is generally required for the hydrophobic interactions, was not essential for the hydrophobic immobilization. The enzyme in free as well as immobilized f orm was optimally active between pH 7.0 and 9.0. The immobilized preparatio n could be reused in a batch process for the conversion Of D-amino acids to cx-keto acids. When the activity of the preparation dropped below practica l limits, the gel could be regenerated by water wash and recharged with fre sh crude extract from yeast.