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Literature summary for 2.5.1.153 extracted from

  • Nakano, C.; Ootsuka, T.; Takayama, K.; Mitsui, T.; Sato, T.; Hoshino, T.
    Characterization of the Rv3378c gene product, a new diterpene synthase for producing tuberculosinol and (13R,S)-isotuberculosinol (nosyberkol), from the Mycobacterium tuberculosis H37Rv genome (2011), Biosci. Biotechnol. Biochem., 75, 75-81.
    View publication on PubMed
Search for more literature for 2.5.1.153

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
D81N 24% activity compared to the wild type enzyme Mycobacterium tuberculosis
D81N/D82N inactive Mycobacterium tuberculosis
D81N/D85N inactive Mycobacterium tuberculosis
D82N 21% activity compared to the wild type enzyme Mycobacterium tuberculosis
D82N/D85N inactive Mycobacterium tuberculosis
D85N 37% activity compared to the wild type enzyme Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
Cu2+ 17% activity at 1 mM Mycobacterium tuberculosis
Fe2+ 20% activity at 1 mM Mycobacterium tuberculosis
Mn2+ 13% activity at 1 mM Mycobacterium tuberculosis
Ni2+ 35% activity at 1 mM Mycobacterium tuberculosis
Zn2+ 10% activity at 1 mM Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0254
-
 tuberculosinyl diphosphate mutant enzyme D82N, at pH 7.0, 45°C Mycobacterium tuberculosis
0.0271
-
 tuberculosinyl diphosphate wild type enzyme, at pH 7.0, 45°C Mycobacterium tuberculosis
0.0273
-
 tuberculosinyl diphosphate mutant enzyme D85N, at pH 7.0, 45°C Mycobacterium tuberculosis
0.0396
-
 tuberculosinyl diphosphate mutant enzyme D81N, at pH 7.0, 45°C Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ essential for activity, maximum activity at 1 mM Mg2+ Mycobacterium tuberculosis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
30000
-
 2 * 30000, SDS-PAGE Mycobacterium tuberculosis
60000
-
 gel filtration Mycobacterium tuberculosis

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
-
-
Mycobacterium tuberculosis H37Rv
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography, amylase column chromatography, and G3000SWXL gel filtration Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 tuberculosinyl diphosphate + 2 H2O
-
 Mycobacterium tuberculosis (13R)-isotuberculosinol + (13S)-isotuberculosinol + 2 diphosphate isotuberculosinol is also named nosyberkol, the product ratio of (13R)-isotuberculosinol and (13S)-isotuberculosinol is 1:3 ?
2 tuberculosinyl diphosphate + 2 H2O
-
 Mycobacterium tuberculosis H37Rv (13R)-isotuberculosinol + (13S)-isotuberculosinol + 2 diphosphate isotuberculosinol is also named nosyberkol, the product ratio of (13R)-isotuberculosinol and (13S)-isotuberculosinol is 1:3 ?
additional information no activity with geranyl diphosphate, farnesyl diphosphate or geranylgeranyl diphosphate Mycobacterium tuberculosis ?
-
 ?
additional information no activity with geranyl diphosphate, farnesyl diphosphate or geranylgeranyl diphosphate Mycobacterium tuberculosis H37Rv ?
-
 ?
tuberculosinyl diphosphate + H2O
-
 Mycobacterium tuberculosis tuberculosinol + diphosphate
-
 ?
tuberculosinyl diphosphate + H2O
-
 Mycobacterium tuberculosis H37Rv tuberculosinol + diphosphate
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 30000, SDS-PAGE Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
Rv3378c protein
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
-
 Mycobacterium tuberculosis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
35 45 no activity at temperatures higher than 50°C Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0000998
-
 tuberculosinyl diphosphate mutant enzyme D82N, at pH 7.0, 45°C Mycobacterium tuberculosis
0.00018
-
 tuberculosinyl diphosphate mutant enzyme D81N, at pH 7.0, 45°C Mycobacterium tuberculosis
0.000192
-
 tuberculosinyl diphosphate mutant enzyme D85N, at pH 7.0, 45°C Mycobacterium tuberculosis
0.000517
-
 tuberculosinyl diphosphate wild type enzyme, at pH 7.0, 45°C Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
 Mycobacterium tuberculosis

pH Range

pH Minimum pH Maximum Comment Organism
5.5 9
-
 Mycobacterium tuberculosis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.00398
-
 tuberculosinyl diphosphate mutant enzyme D82N, at pH 7.0, 45°C Mycobacterium tuberculosis
0.0045
-
 tuberculosinyl diphosphate mutant enzyme D81N, at pH 7.0, 45°C Mycobacterium tuberculosis
0.007
-
 tuberculosinyl diphosphate mutant enzyme D85N, at pH 7.0, 45°C Mycobacterium tuberculosis
0.0191
-
 tuberculosinyl diphosphate wild type enzyme, at pH 7.0, 45°C Mycobacterium tuberculosis