Improved purification of Aspergillus oryzae 1,2-alpha-mannosidase by using mannan gel affinity chromatography.
In order to facilitate the purification of 1,2-alpha-mannosidase from an enzyme product of Aspergillus oryzae, we have devised a rapid and simple procedure. A partially purified enzyme preparation obtained from the A. oryzae enzyme product, by means of ammonium sulfate fractionation followed by CM-Sephadex C-50 chromatography, was subjected to affinity chromatography with baker's yeast mannan gel as an adsorbent. 1,2-alpha-Mannosidase was retarded and well separated from the major protein peak on the affinity column. After a second affinity chromatography under the same conditions, 1,2-alpha-mannosidase was finally purified 7,500-fold with a 22.9% yield. The enzyme preparation thus obtained was quite suitable for the structural analysis of glycoconjugates.
K
K
Tanimoto
T
T
Nishimoto
F
F
Saitoh
H
H
Yamaguchi
D001230:Aspergillus / Q000201:enzymology*; D001236:Aspergillus oryzae / Q000201:enzymology*; D002846:Chromatography, Affinity / Q000379:methods*; D008351:Mannans; D008361:Mannosidases / Q000302:isolation & purification*; D043323:alpha-Mannosidase
D008351:Mannans; D008361:Mannosidases; D043323:alpha-Mannosidase