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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
1988-6-2
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pubmed:abstractText |
Carboxymethyl dextrans (CM-Ds) were used on an HPLC ion-exchange column to obtain significantly enriched alkaline phosphatase (EC 3.1.3.1) from a sample of Escherichia coli periplasmic space proteins without significant loss of enzymatic activity. The ability of CM-Ds to separate alkaline phosphatase even when the column was 80-85% saturated with protein demonstrates the potential for high column capacity using CM-Ds. In addition, the fractions containing alkaline phosphatase and CM-Ds were reapplied to the same ion-exchange column under different buffer conditions and purified to homogeneity by salt gradient elution chromatography, thus demonstrating the compatibility of CM-Ds with the latter chromatographic method. The two-step chromatographic procedure yielded enzyme of purity comparable to that of electrophoretically purified E. coli alkaline phosphatase obtained commercially. These studies demonstrate that HPLC displacement chromatography is a mild procedure which allows rapid, quantitative purification of an enzyme. Scaling up with larger columns should allow purification of enzymes of a commercial basis.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
0003-2697
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
168
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
25-30
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pubmed:dateRevised |
2000-12-18
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pubmed:meshHeading |
pubmed-meshheading:2452588-Alkaline Phosphatase,
pubmed-meshheading:2452588-Chromatography, High Pressure Liquid,
pubmed-meshheading:2452588-Chromatography, Ion Exchange,
pubmed-meshheading:2452588-Dextrans,
pubmed-meshheading:2452588-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:2452588-Escherichia coli,
pubmed-meshheading:2452588-Nucleic Acids
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pubmed:year |
1988
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pubmed:articleTitle |
Purification of Escherichia coli alkaline phosphatase on an ion-exchange high-performance liquid chromatographic column using carboxymethyl dextrans.
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pubmed:affiliation |
Department of Laboratory Medicine, Yale University School of Medicine, New Haven, Connecticut 06510.
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pubmed:publicationType |
Journal Article
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