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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
3
|
| pubmed:dateCreated |
1996-12-3
|
| pubmed:abstractText |
A cDNA encoding soybean alpha-D-galactosidase [E.C. 3.2.1.22] was obtained by screening a soybean library with Phaseolus alpha-D-galactosidase cDNA. The Glycine max alpha-D-galactosidase cDNA is 1.75 kb long and contains untranslated 5' and 3' sequences. The deduced amino acid sequence of the soybean gene has a high degree of homology with other eucaryotic alpha-D-galactosidases. Recombinant alpha-D-galactosidase (rGal) was expressed in Pichia pastoris and purified by affinity chromatography. Purified rGal was homogeneous as judged by SDS-PAGE analysis with the relative molecular mass under reducing conditions of 39.8, and under nonreducing conditions 38.0 kDa. The expressed protein contained the sequence NGLGHTPPMG at the N-terminus, corresponding to the deduced amino acid sequence of the soybean gene. The relative native molecular mass by Sephacryl S-200 chromatography was determined to be 33.1 kDa. The specific activity was 295.6 mumoles of PNP-alpha-D-galactopyranoside hydrolyzed per mg pure rGal per min. rGal was highly specific for alpha-D-galactosyl residues. No detectable hemagglutinin or protease activity was present in the preparations. Furthermore, rGal was active against the blood group B antigen in native human erythrocyte cell suspension assays. The only detectable erythrocyte phenotypic change was loss of the B and P1 epitopes. Consequently, recombinant Glycine max alpha-D-galactosidase may have useful biotechnical applications in the potential mass production of universally transfusable type O erythrocytes by enzymatic conversion.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/ABO Blood-Group System,
http://linkedlifedata.com/resource/pubmed/chemical/Galactosides,
http://linkedlifedata.com/resource/pubmed/chemical/Plant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/alpha-Galactosidase
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Jun
|
| pubmed:issn |
1039-9712
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
39
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
471-85
|
| pubmed:dateRevised |
2004-11-17
|
| pubmed:meshHeading |
pubmed-meshheading:8828798-ABO Blood-Group System,
pubmed-meshheading:8828798-Amino Acid Sequence,
pubmed-meshheading:8828798-Base Sequence,
pubmed-meshheading:8828798-Chromatography, Affinity,
pubmed-meshheading:8828798-Chromatography, Gel,
pubmed-meshheading:8828798-Cloning, Molecular,
pubmed-meshheading:8828798-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:8828798-Erythrocytes,
pubmed-meshheading:8828798-Galactosides,
pubmed-meshheading:8828798-Humans,
pubmed-meshheading:8828798-Hydrogen-Ion Concentration,
pubmed-meshheading:8828798-Molecular Sequence Data,
pubmed-meshheading:8828798-Molecular Weight,
pubmed-meshheading:8828798-Osmolar Concentration,
pubmed-meshheading:8828798-Pichia,
pubmed-meshheading:8828798-Plant Proteins,
pubmed-meshheading:8828798-Recombinant Proteins,
pubmed-meshheading:8828798-Sequence Analysis,
pubmed-meshheading:8828798-Sequence Homology, Amino Acid,
pubmed-meshheading:8828798-Soybeans,
pubmed-meshheading:8828798-Substrate Specificity,
pubmed-meshheading:8828798-alpha-Galactosidase
|
| pubmed:year |
1996
|
| pubmed:articleTitle |
Cloning, expression and characterization of a blood group B active recombinant alpha-D-galactosidase from soybean (Glycine max).
|
| pubmed:affiliation |
Division of Biological Sciences, University of Missouri, Columbia 65212, USA.
|
| pubmed:publicationType |
Journal Article
|