| dc.contributor.author |
Labrou, NE |
en |
| dc.date.accessioned |
2014-06-06T06:44:19Z |
|
| dc.date.available |
2014-06-06T06:44:19Z |
|
| dc.date.issued |
2000 |
en |
| dc.identifier.issn |
0923179X |
en |
| dc.identifier.uri |
http://dx.doi.org/10.1023/A:1008131320571 |
en |
| dc.identifier.uri |
http://62.217.125.90/xmlui/handle/123456789/1800 |
|
| dc.subject |
Candida boidinii |
en |
| dc.subject |
Dye-ligand affinity chromatography |
en |
| dc.subject |
Enzyme purification |
en |
| dc.subject |
Formate dehydrogenase |
en |
| dc.subject.other |
affinity chromatography |
en |
| dc.subject.other |
analytical procedures and techniques |
en |
| dc.subject.other |
anion exchange chromatography |
en |
| dc.subject.other |
Candida boidinii |
en |
| dc.subject.other |
elution |
en |
| dc.subject.other |
enzyme activity |
en |
| dc.subject.other |
enzyme purification |
en |
| dc.subject.other |
formate dehydrogenase |
en |
| dc.subject.other |
ion exchange chromatography |
en |
| dc.subject.other |
molecular weight |
en |
| dc.subject.other |
Acrylics |
en |
| dc.subject.other |
Affinity chromatography |
en |
| dc.subject.other |
Amino acids |
en |
| dc.subject.other |
Electrophoresis |
en |
| dc.subject.other |
Filtration |
en |
| dc.subject.other |
Fungi |
en |
| dc.subject.other |
High performance liquid chromatography |
en |
| dc.subject.other |
Ion exchange |
en |
| dc.subject.other |
Molecular structure |
en |
| dc.subject.other |
Molecular weight |
en |
| dc.subject.other |
Purification |
en |
| dc.subject.other |
Sodium compounds |
en |
| dc.subject.other |
Anion exchange chromatography |
en |
| dc.subject.other |
Elution |
en |
| dc.subject.other |
Formate dehydrogenase |
en |
| dc.subject.other |
Gel electrophoresis |
en |
| dc.subject.other |
Ligands |
en |
| dc.subject.other |
Polyacrylamide |
en |
| dc.subject.other |
Sodium dodecyl sulfate |
en |
| dc.subject.other |
Enzymes |
en |
| dc.subject.other |
Amino Acid Sequence |
en |
| dc.subject.other |
Candida |
en |
| dc.subject.other |
Chromatography, Ion Exchange |
en |
| dc.subject.other |
Formate Dehydrogenases |
en |
| dc.subject.other |
Hydrogen-Ion Concentration |
en |
| dc.subject.other |
Molecular Sequence Data |
en |
| dc.subject.other |
Molecular Weight |
en |
| dc.subject.other |
Candida boidinii |
en |
| dc.subject.other |
Fungi |
en |
| dc.title |
Improved purification of Candida boidinii formate dehydrogenase |
en |
| heal.type |
journalArticle |
en |
| heal.identifier.primary |
10.1023/A:1008131320571 |
en |
| heal.publicationDate |
2000 |
en |
| heal.abstract |
Formate dehydrogenase (FDH, EC 1.2.1.2) from Candida boidinii was purified to homogeneity. The two step procedure comprised anion exchange chromatography (2.9-fold purification, 85% step yield, elution with 35 mM KCl), followed by dye-ligand affinity chromatography on immobilized Cibacron Blue 3GA (1.4-fold purification, 75% step yield, elution with 0.15 mM NAD+/2 mM Na2SO3). The procedure afforded FDH at 63.8% overall yield and a specific activity of 7.2 units/mg. The purity of the final FDH preparation was evaluated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), high performance gel filtration liquid chromatography (gfHPLC) and N-terminal amino acid sequencing. The analytical techniques showed the presence of a single polypeptide chain that corresponds to the molecular weight of 41 kDa (as determined by SDS-PAGE) and 81 kDa (as determined by gfHPLC).Formate dehydrogenase (FDH, EC 1.2.1.2) from Candida boidinii was purified to homogeneity. The two step procedure comprised anion exchange chromatography (2.9-fold purification, 85% step yield, elution with 35 mM KCl), followed by dye-ligand affinity chromatography on immobilized Cibacron Blue 3GA (1.4-fold purification, 75% step yield, elution with 0.15 mM NAD+/2 mM Na2SO3). The procedure afforded FDH at 63.8% overall yield and a specific activity of 7.2 units/mg. The purity of the final FDH preparation was evaluated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), high performance gel filtration liquid chromatography (gfHPLC) and N-terminal amino acid sequencing. The analytical techniques showed the presence of a single polypeptide chain that corresponds to the molecular weight of 41 kDa (as determined by SDS-PAGE) and 81 kDa (as determined by gfHPLC). |
en |
| heal.publisher |
Kluwer Academic Publishers, Dordrecht, Netherlands |
en |
| heal.journalName |
Bioseparation |
en |
| dc.identifier.issue |
2 |
en |
| dc.identifier.volume |
9 |
en |
| dc.identifier.doi |
10.1023/A:1008131320571 |
en |
| dc.identifier.spage |
99 |
en |
| dc.identifier.epage |
104 |
en |