Open AccessPurification and Characterization of a β‐ N ‐Acetylglucosaminidase from Octopus vulgaris
Author(s) -
CECCARINI Costante,
D'ANIELLO Antimo,
CACACE Marcello G.,
ATKINSON Paul H.
Publication year - 1983
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1983.tb07385.x
Subject(s) - exoglycosidase , tetramer , isoelectric focusing , chemistry , chromatography , gel electrophoresis , isoelectric point , sephadex , polyacrylamide gel electrophoresis , gel permeation chromatography , biochemistry , enzyme , glycan , glycoprotein , organic chemistry , polymer
We have purified a β‐ N ‐acetylglucosaminidase from the hepatopancreas of the octopus which we have called βI. The enzyme was homogeneous as judged by Sephadex column chromatography, isoelectric focusing, non‐denaturing gel electrophoresis at two different pH and with sodium dodecyl sulphate/polacrylamide gel electrophoresis. The native portein has an apparent molecular weight of 120000 and we can conclude that it is a tetramer made up of two α and two β subunits with apparent M r of 27000 and 34000, respectively. Using NMR spectroscopy we have examined the specificity of β and have established that the enzyme hydrolyses the β1,4 linkage of N ‐acetylglucosamine but at only a specific site of the substrates used, two glycopeptides isolated from linkage of N ‐acetylglucosamine but at only a specfic site of the substrates used, two glycopeptides isolated from ovalbumin. To our knowledge this is the first known exoglycosidase which has both linkage and site specificty.