Preparation of a Universal Blood Donor Type.
Annual rept. no. 4, 1 Dec 78-30 Nov 79,
MICHIGAN UNIV ANN ARBOR SIMPSON MEMORIAL INST
Pagination or Media Count:
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 oH cross-reactivity. SDS-PAGE analysis results in the detection of two bands of activity when tested with p-nitrophenyl-alpha-N-acetylgalactosaminide, with Re0.3 and 0.44, in the relative amounts of 12. However, only the 0.44 band shows enzymatic activity with ARBC, 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 Re0.28.
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