| http://purl.uniprot.org/citations/8619622 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/8619622 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/8619622 | http://www.w3.org/2000/01/rdf-schema#comment | "Alpha-Galactosidase (alpha-GAL) purified from green coffee bean cleaves the terminal galactose residues from the surface of group B erythrocytes, thereby converting these cells serologically to group O cells. Such enzymatically converted red cells have been transfused into group A and O recipients as part of the first phase of FDA-approved clinical trials. Recently we expressed the recombinant alpha-GAL (r)alpha-GAL) in large quantities in a methylotrophic yeast strain Pichia pastoris and purified the protein to apparent homogeneity by chromatography on a macro prep S50 column. Purified (r)alpha-GAL, migrating as a single band of 41 kDa on a SDS-PAGE, appears to be identical to its native counterpart in specific activity (32 U/mg) and kinetic parameters (K(m) =0.363 mM and V(max) = 46.9 U/mg). Both enzymes demonstrate the same pH profile in the pH range from 2 to 9, with an optimal pH at 6.4 when tested with the substrate p-nitrophenol-alpha-D-galactopyranoside. Furthermore, as with its native counterpart, (r)alpha-GAL specifically cleaves alpha-linked terminal galactose residues from group B red cells without affecting other major antigens on the red cell surface. In addition, we developed a method for using RT-PCR to detect possible DNA contamination in the purified protein preparation, which is one of the concerns for in vivo studies. Thus, with a simple procedure for over-expression and purification of (r)alpha-GAL from P. pastoris culture, one can readily obtain the enzyme needed for large-scale sero-conversion of red cells."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.org/dc/terms/identifier | "doi:10.1006/abbi.1996.0129"xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.org/dc/terms/identifier | "doi:10.1006/abbi.1996.0129"xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Zhang Z."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Zhang Z."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Zhu A."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Zhu A."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Goldstein J."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Goldstein J."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Hurst R."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Hurst R."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Leng L."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Leng L."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Lenny L."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Lenny L."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Monahan C."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/author | "Monahan C."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/date | "1996"xsd:gYear |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/date | "1996"xsd:gYear |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/name | "Arch. Biochem. Biophys."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/name | "Arch. Biochem. Biophys."xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/pages | "324-329"xsd:string |
| http://purl.uniprot.org/citations/8619622 | http://purl.uniprot.org/core/pages | "324-329"xsd:string |