Purification of nodule-specific uricase from soybean by arginine-sepharose affinity chromatography.
A method to purify uricase from soybean root nodules is described. The separation uses a single affinity chromatography step on Arginine-Sepharose, which was constructed by coupling L-Arginine to Activated CH-Sepharose 4B. Crude extracts were loaded onto small columns of Arginine-Sepharose and a significant retardation of uricase was observed. With a re-run of the fraction containing maximal uricase activity on the same column highly purified enzyme was obtained. Analysis by SDS-polyacrylamide gel electrophoresis revealed two protein bands. Analytical isoelectric focusing showed two isoforms of uricase, one dominating with pI 9.0 and a minor with pI 7.0.
Larsen
K
K
Department of Molecular Biology and Plant Physiology, University of Aarhus, Denmark.
D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D013025:Glycine max / Q000201:enzymology*; D014503:Urate Oxidase / Q000302:isolation & purification* / Q000378:metabolism
D014503:Urate Oxidase