Kanamycin nucleotidyltransferase from Staphylococcus aureus

Enzyme Description

Species

Extremophile

EC Number

2.7.7.47 ↗

Sequence

Length: 253 amino acids

MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFFSILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF

Hans H. Liao et al. (1993) — Thermostable mutants of kanamycin nucleotidyltransferase are also more stable to proteinase K, urea, detergents, and water-miscible organic solvents

Enzyme and Microbial Technology · doi:10.1016/0141-0229(93)90151-Q ↗ · Activity - Classical Activity + Stability - Incubation

48 measurements ▼

Database ID

UniProt: P05057 ↗

Sequence Annotation

Inferred - from protein name

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (48 measurements)

48 measurements — page 3 of 3

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
D80Y	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	25%	100.0	4.0	80.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin , Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	30%	100.0	0.0	20.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin , Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	18%	100.0	38.0	91.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin , Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	24%	100.0	7.0	64.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin , Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	30%	100.0	2.0	22.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin , Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	15%	100.0	80.0	98.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin , Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	20%	100.0	57.0	98.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin , Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	25%	100.0	4.0	83.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin , Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent

WT	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	4%	100.0	—	101.6	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
WT	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	8%	100.0	—	98.1	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
WT	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	12%	100.0	—	33.2	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
WT	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	16%	100.0	—	4.3	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
WT	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	20%	100.0	—	0.8	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	24%	100.0	—	7.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	18%	100.0	—	38.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	Inferred - from protein name (2 mutations)	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	30%	100.0	—	2.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	15%	100.0	—	80.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	20%	100.0	—	57.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	25%	100.0	—	4.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
WT	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	30%	100.0	—	0.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	20%	100.0	0.8	14.5	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	20%	100.0	0.8	69.2	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	4%	100.0	101.6	120.8	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	8%	100.0	98.1	98.1	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	12%	100.0	33.2	87.2	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	16%	100.0	4.3	43.4	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	4%	100.0	101.6	106.3	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	8%	100.0	98.1	102.8	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	12%	100.0	33.2	94.6	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	16%	100.0	4.3	59.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	20%	100.0	0.8	32.4	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	4%	100.0	101.6	130.6	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	8%	100.0	98.1	129.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	12%	100.0	33.2	116.1	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity - Classical	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Tetrahydrofuran (THF)	16%	100.0	4.3	97.7	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	15%	100.0	80.0	107.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	18%	100.0	38.0	87.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	24%	100.0	7.0	27.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	30%	100.0	2.0	5.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	15%	100.0	80.0	107.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	20%	100.0	57.0	102.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	25%	100.0	4.0	44.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	30%	100.0	0.0	7.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	18%	100.0	38.0	90.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	24%	100.0	7.0	28.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	30%	100.0	2.0	5.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	30%	100.0	0.0	61.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	20%	100.0	57.0	106.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	25%	100.0	4.0	80.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	30%	100.0	0.0	20.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	18%	100.0	38.0	91.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	24%	100.0	7.0	64.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Dimethylformamide (DMF)	30%	100.0	2.0	22.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	15%	100.0	80.0	98.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	20%	100.0	57.0	98.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent
D80Y/T130K	Activity + Stability - Incubation	Activity after incubation (15 minutes at 22-24°C), measured by radioactivity assay (radioactivity measurement of [³H]AMP-kanamycin binding to phosphocellulose paper) in the presence of organic solvent	Ethanol	25%	100.0	4.0	83.0	%	?	?	Incubation: 22-24°C, Assay: 37°C	? mM AMP-kanamycin, ? mM ATP (Adenosine triphosphate)	AMP–kanamycin, Pyrophosphate	—	—	Non-incubated control (in %), assay in the presence of organic solvent \| Clear about assay in the presence of organic solvent

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Mutations in this dataset (4)

WT D80Y D80Y/T130K T130K

Visualization : Activity — Classical

One bar per measurement. Colour = solvent, shade = solvent volume. — — — Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Visualization : Activity + Stability — Incubation

One bar per measurement. Colour = solvent, shade = solvent volume. — — — Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Structure

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Kanamycin nucleotidyltransferase from Staphylococcus aureus

Enzyme Description

Sequence

Experimental Data (48 measurements)

Visualization : Activity — Classical

Mutation Effect

Visualization : Activity + Stability — Incubation

Mutation Effect

Structure

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