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Literature summary for 1.3.1.44 extracted from

  • Hu, K.; Zhao, M.; Zhang, T.; Zha, M.; Zhong, C.; Jiang, Y.; Ding, J.
    Structures of trans-2-enoyl-CoA reductases from Clostridium acetobutylicum and Treponema denticola: insights into the substrate specificity and the catalytic mechanism (2013), Biochem. J., 449, 79-89.
    View publication on PubMed
Search for more literature for 1.3.1.44

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Treponema denticola
expressed in Escherichia coli as a His-tagged fusion protein Clostridium acetobutylicum

Crystallization (Commentary)

Crystallization (Comment) Organism
in complex with NAD+, vapor diffusion method Clostridium acetobutylicum
in complex with NAD+, vapor diffusion method, Treponema denticola
structures of CaTER in apo form at 2.1 A resolution, in complex with NADH at 2.0 A resolution and in complex with NAD+ at 2.7 A resolution Clostridium acetobutylicum
TdTER is crystallized in complex with NAD+ at 3.0 A resolution Treponema denticola

Protein Variants

Protein Variants Comment Organism
F11K kcat slightly decreased compared to wild-type, Km (NADH) decreased compared to wild-type Clostridium acetobutylicum
F11K the mutant shows 198% activity compared to the wild type enzyme Clostridium acetobutylicum
K244A inactive Clostridium acetobutylicum
K244A mutant shows no activity Clostridium acetobutylicum
K245A kcat slightly decreased compared to wild-type, Km (NADH) increased compared to wild-type Clostridium acetobutylicum
K245A the mutant shows 67 activity compared to the wild type enzyme Clostridium acetobutylicum
Y225A kcat highly decreased compared to wild-type, Km (NADH) increased compared to wild-type Clostridium acetobutylicum
Y225A the mutant shows 6.7 activity compared to the wild type enzyme Clostridium acetobutylicum
Y235F inactive Clostridium acetobutylicum
Y235F mutant shows no activity Clostridium acetobutylicum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0316
-
 NADH pH 6.2, 25°C, mutant F11K Clostridium acetobutylicum
0.0316
-
 NADH mutant enzyme F11K, at pH 6.2 and 25°C Clostridium acetobutylicum
0.0697
-
 NADH pH 6.2, 25°C Treponema denticola
0.0697
-
 NADH wild type enzyme, at pH 6.2 and 25°C Treponema denticola
0.1054
-
 NADH pH 6.2, 25°C Clostridium acetobutylicum
0.1054
-
 NADH wild type enzyme, at pH 6.2 and 25°C Clostridium acetobutylicum
0.129
-
 NADH pH 6.2, 25°C, mutant Y225A Clostridium acetobutylicum
0.1299
-
 NADH mutant enzyme Y225A, at pH 6.2 and 25°C Clostridium acetobutylicum
0.132
-
 NADH pH 6.2, 25°C, mutant K245A Clostridium acetobutylicum
0.1323
-
 NADH mutant enzyme K245A, at pH 6.2 and 25°C Clostridium acetobutylicum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
44800
-
-
 Treponema denticola
44800
-
 calculated from cDNA Treponema denticola
45700
-
 calculated from cDNA Clostridium acetobutylicum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
crotonyl-CoA + NADH Clostridium acetobutylicum
-
 butanoyl-CoA + NAD+
-
 ir
crotonyl-CoA + NADH Treponema denticola
-
 butanoyl-CoA + NAD+
-
 ir

Organism

Organism UniProt Comment Textmining
Clostridium acetobutylicum Q97LU2
-
-
Treponema denticola
-
-
-
Treponema denticola Q73Q47
-
-
Treponema denticola ATCC 35405
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Treponema denticola
using Ni-NTA chromatography Clostridium acetobutylicum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
crotonyl-CoA + NADH
-
 Clostridium acetobutylicum butanoyl-CoA + NAD+
-
 ?
crotonyl-CoA + NADH
-
 Clostridium acetobutylicum butanoyl-CoA + NAD+
-
 ir
crotonyl-CoA + NADH
-
 Treponema denticola butanoyl-CoA + NAD+
-
 ir
crotonyl-CoA + NADH high activity toward crotonyl-CoA Treponema denticola butanoyl-CoA + NAD+
-
 ?
crotonyl-CoA + NADH high activity toward crotonyl-CoA Treponema denticola ATCC 35405 butanoyl-CoA + NAD+
-
 ?
crotonyl-CoA + NADPH
-
 Clostridium acetobutylicum butanoyl-CoA + NADP+
-
 ?
hexenoyl-CoA + NADH weak activity toward crotonyl-CoA Treponema denticola ? + NAD+
-
 ?
hexenoyl-CoA + NADH weak activity toward crotonyl-CoA Treponema denticola ATCC 35405 ? + NAD+
-
 ?
additional information no activity with butanoyl-CoA Clostridium acetobutylicum ?
-
 ?
additional information no activity with butanoyl-CoA Treponema denticola ?
-
 ?

Subunits

Subunits Comment Organism
? x * 45700, calculated from amino acid sequence Clostridium acetobutylicum
? x * 44800, calculated from amino acid sequence Treponema denticola
monomer
-
 Treponema denticola
monomer
-
 Clostridium acetobutylicum

Synonyms

Synonyms Comment Organism
CaTER
-
 Clostridium acetobutylicum
tdTer
-
 Treponema denticola
TER
-
 Clostridium acetobutylicum
TER
-
 Treponema denticola
trans-2-enoyl-CoA reductase
-
 Clostridium acetobutylicum
trans-2-enoyl-CoA reductase
-
 Treponema denticola
trans-2-enoyl-CoA reductases
-
 Treponema denticola
trans-2-enoyl-CoA reductases
-
 Clostridium acetobutylicum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Treponema denticola
25
-
 assay at Clostridium acetobutylicum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3
-
 NADH pH 6.2, 25°C, mutant Y225A Clostridium acetobutylicum
2.3
-
 NADH mutant enzyme Y225A, at pH 6.2 and 25°C Clostridium acetobutylicum
16.5
-
 NADH pH 6.2, 25°C, mutant F11K Clostridium acetobutylicum
16.5
-
 NADH mutant enzyme F11K, at pH 6.2 and 25°C Clostridium acetobutylicum
23.6
-
 NADH pH 6.2, 25°C, mutant K245A Clostridium acetobutylicum
23.6
-
 NADH mutant enzyme K245A, at pH 6.2 and 25°C Clostridium acetobutylicum
28.2
-
 NADH pH 6.2, 25°C, wild-type Clostridium acetobutylicum
28.2
-
 NADH wild type enzyme, at pH 6.2 and 25°C Clostridium acetobutylicum
385.9
-
 NADH pH 6.2, 25°C, wild-type Treponema denticola
385.9
-
 NADH wild type enzyme, at pH 6.2 and 25°C Treponema denticola

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.2
-
 assay at Treponema denticola
6.2
-
 assay at Clostridium acetobutylicum

Cofactor

Cofactor Comment Organism Structure
NADH
-
 Clostridium acetobutylicum
NADH
-
 Treponema denticola
NADH higher activity toward NADH than NADPH Treponema denticola
NADH higher activity toward NADH than NADPH Clostridium acetobutylicum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
18
-
 NADH pH 6.2, 25°C, mutant Y225A Clostridium acetobutylicum
18
-
 NADH mutant enzyme Y225A, at pH 6.2 and 25°C Clostridium acetobutylicum
180
-
 NADH pH 6.2, 25°C, mutant K245A Clostridium acetobutylicum
180
-
 NADH mutant enzyme K245A, at pH 6.2 and 25°C Clostridium acetobutylicum
270
-
 NADH pH 6.2, 25°C, wild-type Clostridium acetobutylicum
270
-
 NADH wild type enzyme, at pH 6.2 and 25°C Clostridium acetobutylicum
520
-
 NADH pH 6.2, 25°C, mutant F11K Clostridium acetobutylicum
520
-
 NADH mutant enzyme F11K, at pH 6.2 and 25°C Clostridium acetobutylicum
5500
-
 NADH pH 6.2, 25°C, wild-type Treponema denticola
5500
-
 NADH wild type enzyme, at pH 6.2 and 25°C Treponema denticola