Immobilized metal ion affinity chromatography in enzyme fractionation.
Ribitol dehydrogenase from Mycobacterium butyricum and alpha-mannosidase from Lupinus luteus seedlings were fractionated by the immobilized metal ion (Cu2+ or Zn2+) affinity chromatography (IMAC) on iminodiacetic acid coupled to Sepharose 6B. In a single step, ribitol dehydrogenase was purified 10-12 fold with the recovery above 80% when using Zn(2+)-Sepharose 6B as the sorbent and decreasing linear gradient of pH from 7 to 4. In the same conditions purification of alpha-mannosidase was less effective (2-3 fold, recovery 60-70%).
Byra
A
A
Department of Biochemistry and Drug Metabolism, Medical School Lublin, Poland.
Dworniczak
K
K
Szumiło
T
T
D002846:Chromatography, Affinity / Q000379:methods; D003300:Copper; D008361:Mannosidases / Q000302:isolation & purification*; D008670:Metals; D009161:Mycobacterium / Q000201:enzymology; D010944:Plants / Q000201:enzymology; D013401:Sugar Alcohol Dehydrogenases / Q000302:isolation & purification*; D015032:Zinc; D043323:alpha-Mannosidase
D008670:Metals; D003300:Copper; D013401:Sugar Alcohol Dehydrogenases; C022529:ribitol 2-dehydrogenase; D008361:Mannosidases; D043323:alpha-Mannosidase; D015032:Zinc