Preparation of a Universal Blood Donor Type.
Abstract
The A-zyme from clostridium perfringens has been purified over 8,000 fold with a yield of 17%. The contaminating enzymes: sialidase, beta-galactosidase and beta-N-acetylglucosaminidase, that have persistently co-purified with it, have new been reduced to 1 - 2% of the level of A-zyme activity. Treatment of various mucins with the purified enzyme released predominantly N-acetylgalactosamine from A-active glycoproteins, and only trace amounts of n-acetylglucosamine. This action is accompanied by a loss of A activity and the development of (o)H cross-reactivity. SDS-PAGE analysis results in the detection of two bands of activity when tested with p-nitrophenyl-alpha-N-acetylgalactosaminide, with Re=0.3 and 0.44, in the relative amounts of 1:2. However, only the 0.44 band shows enzymatic activity with A+RBC, glycoproteins and terminal non-reducing N-acetylgalactosamine containing oligosaccharides derived therefrom. The B-zyme has now been purified 2,500 fold from clostridium sporogenes, Maebashi, with a recovery of 4%. No other glycosidase was detected as a contaminant. Disc gel electrophoresis demonstrated the presence of several protein staining bands, with the alpha-galactosidase activity restricted to essentially one band at Re=0.28.
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 10, 1979
- Accession Number
- ADA080383
Entities
People
- David Aminoff
Organizations
- University of Michigan