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Literature summary for 5.4.99.16 extracted from

  • Wang, Y.; Zhang, J.; Wang, W.; Liu, Y.; Xing, L.; Li, M.
    Effects of the N-terminal and C-terminal domains of Meiothermus ruber CBS-01 trehalose synthase on thermostability and activity (2012), Extremophiles, 16, 377-385.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene treS, sequence comparison, expression of His6-tagged wild-type enzyme and chimeric mutant in Escherichia coli strain Rosetta-gami (DE3), subcloning in Escherichia coli strain DH5alpha Thermus thermophilus
gene treS, sequence comparison, expression of His6-tagged wild-type enzyme and the chimeric mutant in Escherichia coli strain Rosetta-gami (DE3), subcloning in Escherichia coli strain DH5alpha Meiothermus ruber

Protein Variants

Protein Variants Comment Organism
additional information construction of a chimeric enzyme mutant consisting of the C-terminus from Meiothermus ruber trehalose synthase and the N-terminus from Thermus thermophilus trehalose synthase, TSMrTt , and a second with N-terminus from Meiothermus ruber trehalose synthase and C-terminus from Thermus thermophilus trehalose synthase, TSTtMr Meiothermus ruber
additional information construction of a chimeric enzyme mutant consisting of the C-terminus from Meiothermus ruber trehalose synthase and the N-terminus from Thermus thermophilus trehalose synthase, TSMrTt , and a second with N-terminus from Meiothermus ruber trehalose synthase and C-terminus from Thermus thermophilus trehalose synthase, TSTtMr Thermus thermophilus
R392A site-directed mutagenesis, the mutant shows a sharp decrease in activity compared to the wild-type enzyme Meiothermus ruber
R392F site-directed mutagenesis, the mutation leads to a complete loss in activity Meiothermus ruber

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
57.5
-
maltose pH 6.5, 30°C, recombinant mutant TSTtMr Meiothermus ruber
57.5
-
maltose pH 6.5, 30°C, recombinant mutant TSTtMr Thermus thermophilus
69.6
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant mutant TSMrTt Meiothermus ruber
69.6
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant mutant TSMrTt Thermus thermophilus
79.2
-
maltose pH 6.5, 30°C, recombinant mutant TSMrTt Meiothermus ruber
79.2
-
maltose pH 6.5, 30°C, recombinant mutant TSMrTt Thermus thermophilus
82.2
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant wild-type enzyme Thermus thermophilus
97.4
-
maltose pH 6.5, 30°C, recombinant wild-type enzyme Thermus thermophilus
98.9
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant wild-type enzyme Meiothermus ruber
126
-
maltose pH 6.5, 30°C, recombinant wild-type enzyme Meiothermus ruber
154
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant mutant TSTtMr Meiothermus ruber
154
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant mutant TSTtMr Thermus thermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
maltose Meiothermus ruber
-
alpha,alpha-trehalose
-
r
maltose Thermus thermophilus
-
alpha,alpha-trehalose
-
r
maltose Meiothermus ruber CBS-01
-
alpha,alpha-trehalose
-
r
maltose Thermus thermophilus HB8 / ATCC 27634 / DSM 579
-
alpha,alpha-trehalose
-
r

Organism

Organism UniProt Comment Textmining
Meiothermus ruber B1PK99 gene treS
-
Meiothermus ruber CBS-01 B1PK99 gene treS
-
Thermus thermophilus Q5SL15 gene treS
-
Thermus thermophilus HB8 / ATCC 27634 / DSM 579 Q5SL15 gene treS
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged wild-type and chimeric mutant enzymes from Escherichia coli strain Rosetta-gami (DE3) by nickel affinity chromatography Meiothermus ruber
recombinant His6-tagged wild-type and chimeric mutant enzymes from Escherichia coli strain Rosetta-gami (DE3) by nickel affinity chromatography Thermus thermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
alpha,alpha-trehalose
-
Meiothermus ruber maltose
-
r
alpha,alpha-trehalose
-
Thermus thermophilus maltose
-
?
alpha,alpha-trehalose
-
Meiothermus ruber CBS-01 maltose
-
r
alpha,alpha-trehalose
-
Thermus thermophilus HB8 / ATCC 27634 / DSM 579 maltose
-
?
maltose
-
Meiothermus ruber alpha,alpha-trehalose
-
r
maltose
-
Thermus thermophilus alpha,alpha-trehalose
-
r
maltose
-
Meiothermus ruber CBS-01 alpha,alpha-trehalose
-
r
maltose
-
Thermus thermophilus HB8 / ATCC 27634 / DSM 579 alpha,alpha-trehalose
-
r

Subunits

Subunits Comment Organism
More the isolated N-terminal domain from Meiothermus ruber is not active. The secondary structure of the isolated N-terminal domain undergoes a greater change than that of the isolated C-terminus, three-dimensional structure analysis and modeling, overview Thermus thermophilus
More the isolated N-terminal domain from Meiothermus ruber is not active. The secondary structure of the isolated N-terminal domain undergoes a greater change than that of the isolated C-terminus, three-dimensional structure enzyme structure analysis and molecular modeling, overview Meiothermus ruber

Synonyms

Synonyms Comment Organism
Trehalose synthase
-
Meiothermus ruber
Trehalose synthase
-
Thermus thermophilus
TreS
-
Meiothermus ruber
TreS
-
Thermus thermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
recombinant wild-type enzyme and mutant TSMrTt Meiothermus ruber
60
-
recombinant wild-type enzyme and mutant TSTtMr Thermus thermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
70
-
half-lives of purified wild-type enzyme: 48.5 min, purified mutant TSMrTt : 47.6 min, and purified mutant TSTtMr: over 480 min, mutant R392A: 42.2 min Meiothermus ruber
70
-
half-lives of purified wild-type enzyme: over 480 min, purified mutant TSMrTt : 47.6 min, and purified mutant TSTtMr: over 480 min Thermus thermophilus
75
-
half-lives of purified wild-type enzyme: 15.8 min, purified mutant TSMrTt : 17.6 min, and purified mutant TSTtMr: over 480 min, mutant R392A: 12.5 min Meiothermus ruber
75
-
half-lives of purified wild-type enzyme: 475 min, purified mutant TSMrTt : 17.6 min, and purified mutant TSTtMr: over 480 min Thermus thermophilus
80
-
half-lives of purified wild-type enzyme: 233 min, purified mutant TSMrTt : below 5 min, and purified mutant TSTtMr: 27.6 min Thermus thermophilus
80
-
half-lives of purified wild-type enzyme: below 5 min, purified mutant TSMrTt : below 5 min, and purified mutant TSTtMr: 27.6 min, mutant R392A: below 5 min Meiothermus ruber
100
-
purified enzyme, 10 min, inactivation Meiothermus ruber

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
61
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant mutant TSMrTt Meiothermus ruber
68.9
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant wild-type enzyme Meiothermus ruber
110
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant wild-type enzyme Meiothermus ruber
115
-
maltose pH 6.5, 30°C, recombinant mutant TSMrTt Meiothermus ruber
115
-
maltose pH 6.5, 30°C, recombinant mutant TSMrTt Thermus thermophilus
147
-
maltose pH 6.5, 30°C, recombinant wild-type enzyme Meiothermus ruber
152
-
maltose pH 6.5, 30°C, recombinant mutant TSTtMr Meiothermus ruber
152
-
maltose pH 6.5, 30°C, recombinant mutant TSTtMr Thermus thermophilus
202
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant mutant TSTtMr Meiothermus ruber
227
-
maltose pH 6.5, 30°C, recombinant wild-type enzyme Thermus thermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
assay at Meiothermus ruber
6.5
-
assay at Thermus thermophilus

General Information

General Information Comment Organism
additional information enzyme structure analysis and molecular modeling, overview Meiothermus ruber

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.697
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant wild-type enzyme Meiothermus ruber
0.871
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant mutant TSMrTt Meiothermus ruber
0.871
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant mutant TSMrTt Thermus thermophilus
1.16
-
maltose pH 6.5, 30°C, recombinant wild-type enzyme Meiothermus ruber
1.32
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant mutant TSTtMr Meiothermus ruber
1.32
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant mutant TSTtMr Thermus thermophilus
1.33
-
alpha,alpha-trehalose pH 6.5, 30°C, recombinant wild-type enzyme Thermus thermophilus
1.5
-
maltose pH 6.5, 30°C, recombinant mutant TSMrTt Meiothermus ruber
1.5
-
maltose pH 6.5, 30°C, recombinant mutant TSMrTt Thermus thermophilus
2.51
-
maltose pH 6.5, 30°C, recombinant wild-type enzyme Thermus thermophilus
2.7
-
maltose pH 6.5, 30°C, recombinant mutant TSTtMr Meiothermus ruber
2.7
-
maltose pH 6.5, 30°C, recombinant mutant TSTtMr Thermus thermophilus