Expanded bed adsorption (EBA) was successfully applied for the purification of alcohol dehydrogenase (ADH) from unclarified bakers' yeast homogenate using a new dye-iminodiacetic acid (IDA) matrix carrying immobilized metal ions. The existence of cells and cell debris do not have a significant effect on the adsorption of ADH on the matrix. The presence of imidazole in adsorption buffer was found to be an important factor in reducing the level of the adsorbed contaminant proteins. The optimal imidazole concentration was 5 mM. The dynamic binding capacity of the dye-IDA matrix in EBA was found to be 569 U/ml matrix. A chromatography of one-step elution with imidazole was developed, in which ADH was successfully adsorbed in the presence of 5 mM imidazole; subsequently, the column was washed with 5 mM imidazole containing 1 M NaCl and eluted with 150 mM imidazole containing 1.5 M NaCl. The purification factor and ADH recovery were 8.8 and 93.5%, respectively.
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Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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Site | Modification | Modifier | Source | Reference |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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