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Literature summary for 2.7.1.163 extracted from

  • Nakamura, A.; Takakura, Y.; Sugimoto, N.; Takaya, N.; Shiraki, K.; Hoshino, T.
    Enzymatic analysis of a thermostabilized mutant of an Escherichia coli hygromycin B phosphotransferase (2008), Biosci. Biotechnol. Biochem., 72, 2467-2471.
    View publication on PubMed
Search for more literature for 2.7.1.163

Cloned(Commentary)

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

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

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 [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

Metals/Ions Comment Organism Structure
Mg2+ included in assay medium Escherichia coli

Molecular Weight [Da]

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/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
hygromycin B + ATP Escherichia coli
-
 4-O-phosphohygromycin B + ADP
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

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

Source Tissue Comment Organism Textmining
cell culture strain BL23 (DE3) Escherichia coli
-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
hygromycin B + ATP
-
 Escherichia coli 4-O-phosphohygromycin B + ADP
-
 ?

Synonyms

Synonyms Comment Organism
HPH
-
 Escherichia coli
Hph5 thermostable mutant Escherichia coli
hygromycin B phosphotransferase
-
 Escherichia coli

Temperature Optimum [°C]

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 [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Escherichia coli