Cloned (Comment) | Organism |
---|---|
hph5 gene is introduced into Thermus thermophilus on a plasmid of eight copies. HPH and HPH5 are PCR amplified and Escherichia coli strain BL21 (DE3) is transformed with the plasmids and cultivated. | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
A118V | mutant HPH5 with increased thermostability | Escherichia coli |
D20G | mutant HPH5 with increased thermostability | Escherichia coli |
Q226L | mutant HPH5 with increased thermostability | Escherichia coli |
S225P | mutant HPH5 with increased thermostability | Escherichia coli |
T246A | mutant HPH5 with increased thermostability | Escherichia coli |
General Stability | Organism |
---|---|
protein denaturation of HPH at 37.2°C and denaturation of HPH5 at 58.8°C | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.9 | - |
hygromycin B | HPH | Escherichia coli | |
2 | 3 | ATP | HPH and HPH5 | Escherichia coli | |
2.9 | - |
hygromycin B | HPH5 | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | included in assay medium | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
39030 | - |
for HPH5, calculated by amino acid sequence and determined by SDS-PAGE | Escherichia coli |
39100 | - |
for HPH, calculated by amino acid sequence and determined by SDS-PAGE | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
hygromycin B + ATP | Escherichia coli | - |
4-O-phosphohygromycin B + ADP | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
from cell culture by centrifugation and sonication, supernatant is loaded onto a column of chelating sepharose fast flow and then proteins are subjected to gel filtration | Escherichia coli |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
cell culture | strain BL23 (DE3) | Escherichia coli | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
17 | - |
HPH, ph 7.5 at 30°C | Escherichia coli |
21 | - |
HPH5, ph 7.5 at 30°C | Escherichia coli |
35.2 | - |
HPH5, ph 7.5 at 55°C | Escherichia coli |
36.3 | - |
HPH, ph 7.5 at 50°C | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
hygromycin B + ATP | - |
Escherichia coli | 4-O-phosphohygromycin B + ADP | - |
? |
Synonyms | Comment | Organism |
---|---|---|
HPH | - |
Escherichia coli |
Hph5 | thermostable mutant | Escherichia coli |
hygromycin B phosphotransferase | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
optimum temperature for HPH enzyme activity | Escherichia coli |
55 | - |
optimum temperature HPH5 enzyme activity | Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
36 | - |
HPH, thermal stability of enzyme activity | Escherichia coli |
53 | - |
HPH5, thermal stability of enzyme activity | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5 | - |
hygromycin B | HPH | Escherichia coli | |
5.7 | - |
ATP | HPH | Escherichia coli | |
6.4 | - |
ATP | thermostabilized mutant HPH5 | Escherichia coli | |
9.1 | - |
hygromycin B | thermostabilized mutant HPH5 | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli |