A new affinity chromatography using yeast invertase-sepharose 4b for purification of plant endo-β-n-acetylglucosaminidases

For the purification of plant endo-β-N-acetylglucosaminidase, in this report, we introduce a new affinity chromatography using the reduced and carboxymethylated yeast invertase (cm-YI) as a ligand. Two plant endo-β-N-acetylglucosaminidases (endo-LE from tomato fruits (Kimura, Y., et al. Biochim. Biophys. Acta 1381, 27-36 (1998)) and endo-GB from Ginkgo biloba seeds (Kimura, Y., et al. Biosci. Biotechnol. Biochem., 62, 253-261 (1998)) could completely bind to the high-mannose type N-glycans linked to the immobilized yeast invertase and the activities of both enzymes could be recovered by increasing the concentration of NaCl. By using this purification procedure with some other purification procedures, endo-LE could be purified 1,700-fold and endo-GB was purified to apparent homogeneity at 63 kDa as reported previously.