Isolation and characterization of catalase from Penicillium chrysogenum.
Catalase from a crude preparation of Penicillium chrysogenum was isolated in a single chromatographic step by immobilized metal ion affinity chromatography (IMAC) on Cu(II)-Chelating Sepharose Fast Flow. A chromatographically and electrophoretically homogeneous enzyme was obtained in 89% yield. IMAC was found to be superior to ion-exchange, hydrophobic interaction, size-exclusion and concanavalin A affinity chromatography. Analytical and preparative chromatography gave essentially the same chromatograms. Isoelectric point, molecular weight (by ultracentrifugation), amino acid composition, carbohydrate content and subunit organization were determined. The apparent Michaelis-Menten constant, KM, and the azide competitor constant, Ki, were calculated and found to be 59 microM and 6.1 microM, respectively.
Chaga
G S
GS
Institute of Biochemistry, Uppsala University, Sweden.
Medin
A S
AS
Chaga
S G
SG
Porath
J O
JO
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis; D002374:Catalase / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods; D006863:Hydrogen-Ion Concentration; D007477:Ions; D007525:Isoelectric Focusing; D008670:Metals; D008969:Molecular Sequence Data; D010408:Penicillium chrysogenum / Q000201:enzymology*; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D002241:Carbohydrates; D007477:Ions; D008670:Metals; D002374:Catalase