Directed evolution by random PCR mutagenesis of the gene for the aminoglycoside 2''-IIa phosphotransferase generated R92H/D268N and N196D/D268N mutant enzymes, resulting in elevated levels of resistance to amikacin and isepamicin but not to other aminoglycoside antibiotics. Increases in the activities of the mutant phosphotransferases for isepamicin are the result of decreases in K(m) values, more ..
Tested Compounds:
Description:
Title: Mutant APH(2'')-IIa enzymes with increased activity against amikacin and isepamicin.
Abstract: Directed evolution by random PCR mutagenesis of the gene for the aminoglycoside 2''-IIa phosphotransferase generated R92H/D268N and N196D/D268N mutant enzymes, resulting in elevated levels of resistance to amikacin and isepamicin but not to other aminoglycoside antibiotics. Increases in the activities of the mutant phosphotransferases for isepamicin are the result of decreases in K(m) values, while improved catalytic efficiency for amikacin is the result of both a decrease in K(m) values and an increase in turnover of the antibiotic. Enzymes with R92H, D268N, and D268N single amino acid substitutions did not result in elevated MICs for aminoglycosides.
(PMID: 20145089)
Abstract: Directed evolution by random PCR mutagenesis of the gene for the aminoglycoside 2''-IIa phosphotransferase generated R92H/D268N and N196D/D268N mutant enzymes, resulting in elevated levels of resistance to amikacin and isepamicin but not to other aminoglycoside antibiotics. Increases in the activities of the mutant phosphotransferases for isepamicin are the result of decreases in K(m) values, while improved catalytic efficiency for amikacin is the result of both a decrease in K(m) values and an increase in turnover of the antibiotic. Enzymes with R92H, D268N, and D268N single amino acid substitutions did not result in elevated MICs for aminoglycosides.
(PMID: 20145089)
Comment
Putative Target:
ChEMBL Target ID: 22226
Target Type: UNCHECKED
Pref Name: Unchecked
Confidence: Default value - Target unknown or has yet to be assigned
Relationship Type: Default value - Target has yet to be curated
ChEMBL Target ID: 22226
Target Type: UNCHECKED
Pref Name: Unchecked
Confidence: Default value - Target unknown or has yet to be assigned
Relationship Type: Default value - Target has yet to be curated
Categorized Comment
ChEMBL Assay Type: Binding
ChEMBL Assay Data Source: Scientific Literature
ChEMBL Assay Data Source: Scientific Literature
Result Definitions
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| TID | Name | Description | Histogram | Type | Unit | |
|---|---|---|---|---|---|---|
| Outcome | The BioAssay activity outcome | Outcome | ||||
| 1 | Kcat/Km activity comment | Kcat/Km activity comment | String | |||
| 2 | Kcat/Km standard flag | Kcat/Km standard flag |  | Integer | ||
| 3 | Kcat/Km qualifier | Kcat/Km qualifier | String | |||
| 4 | Kcat/Km published value | Kcat/Km published value | | Float | 10'5/M/s | |
| 5 | Kcat/Km standard value | Kcat/Km standard value | | Float | 10'5/M/s | |
| 6 | Kcat/Km activity comment | Kcat/Km activity comment | String | |||
| 7 | Kcat/Km standard flag | Kcat/Km standard flag | | Integer | ||
| 8 | Kcat/Km qualifier | Kcat/Km qualifier | String | |||
| 9 | Kcat/Km published value | Kcat/Km published value | | Float | 10'6/M/s | |
| 10 | Kcat/Km standard value | Kcat/Km standard value | | Float | 10'6/M/s | |
| 11 | Kcat/Km activity comment | Kcat/Km activity comment | String | |||
| 12 | Kcat/Km standard flag | Kcat/Km standard flag | | Integer | ||
| 13 | Kcat/Km qualifier | Kcat/Km qualifier | String | |||
| 14 | Kcat/Km published value | Kcat/Km published value | | Float | 10'7/M/s | |
| 15 | Kcat/Km standard value | Kcat/Km standard value | | Float | 10'7/M/s |
Data Table (Concise)
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