Purification of Rhodotorula gracilis D-amino acid oxidase.
A protocol is presented for preparing Rhodotorula gracilis D-amino acid oxidase in homogeneous form and in high yield in 3 to 4 days. The method takes advantage of (a) cell rupture by alternate freeze-thawing, (b) use of DEAE-Sepharose to bind contaminants, and (c) enzyme binding to a Mono S column. The D-amino acid oxidase isolated by this means has the same spectral and catalytic properties as the enzyme previously obtained, and possesses improved long-term stability.
Pollegioni
L
L
Department of General Physiology and Biochemistry, University of Milan, Italy.
Pilone
M S
MS
D002847:Chromatography, Agarose; D002852:Chromatography, Ion Exchange; D003605:D-Amino-Acid Oxidase / Q000302:isolation & purification*; D005656:Fungal Proteins / Q000302:isolation & purification*; D007700:Kinetics; D012248:Rhodotorula / Q000201:enzymology*
D005656:Fungal Proteins; D003605:D-Amino-Acid Oxidase