Isolation and purification of mitochondrial monoamine oxidase B from bovine liver by chromatofocusing.
Mitochondrial monoamine oxidase B has been isolated and purified from bovine liver. The isolation procedure involves (a) a series of purification steps including isolation of the mitochondria, extraction with non-ionic detergents and chromatofocusing; (b) a novel fluorimetric titration assay. The purified enzyme is isolated following a 8.8 fold purification with an overall yield of 12% and a specific activity of 3460 nM.g-1. A comparison is made with other established procedures. Ion exchange chromatography on DEAE-Cellulose produced a 6.8 fold purification, 6.2% yield and 2383.3 nM.g-1 specific activity while with affinity chromatography on Blue Sepharose CL-6B gave a 3 fold purification, 4.1% yield and a specific activity of 1152.7 nM.g-1.
Daya
S
S
Department of Biochemistry and Microbiology, Rhodes University, Grahamstown Republic of South Africa.
Mia
R B
RB
Whiteley
C G
CG
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D007525:Isoelectric Focusing; D008930:Mitochondria, Liver / Q000201:enzymology*; D008995:Monoamine Oxidase / Q000302:isolation & purification* / Q000378:metabolism; D012701:Serotonin / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D012701:Serotonin; D008995:Monoamine Oxidase