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Literature summary for 1.1.1.71 extracted from

  • Stekhanova, T.N.; Bezsudnova, E.Y.; Mardanov, A.V.; Gumerov, V.M.; Artemova, N.; Kleymenov, S.Y.; Popov, V.O.
    Sodium chloride-induced modulation of the activity and thermal stability of short-chain oxidoreductase from the archaeon Thermococcus sibiricus (2013), Appl. Biochem. Biotechnol., 171, 1877-1889.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a fusion protein with (His)6-tag and 13 amino acid linker at the N-terminal Thermococcus sibiricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.02
-
NADP+ 60°C, pH 10.5, cosubstrate: 2,5-hexandiol Thermococcus sibiricus
0.02
-
NADPH 60°C, pH 7.5, cosubstrate: pyruvaldehyde Thermococcus sibiricus
1.5
-
2,3-Pentanedione 60°C, pH 7.5 Thermococcus sibiricus
13.1
-
(2S,5S)-2,5-hexanediol 60°C, pH 10.5 Thermococcus sibiricus
17.75
-
Pyruvaldehyde 60°C, pH 7.5 Thermococcus sibiricus
168
-
2-propanol 60°C, pH 10.5 Thermococcus sibiricus

Metals/Ions

Metals/Ions Comment Organism Structure
NaCl activity is increased in the presence of NaCl and remains at the elevated level up to 4 M of NaCl. The rates of 2-propanol and 2,5-hexanediol oxidation are increased by more than 2fold after the addition of NaCl up to 1 M to the assay mixture and is retained at the increased level up to 4 M of NaCl Thermococcus sibiricus

Organism

Organism UniProt Comment Textmining
Thermococcus sibiricus C6A190
-
-
Thermococcus sibiricus DSM 12597 C6A190
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(2S,5S)-2,5-hexanediol + NADP+
-
Thermococcus sibiricus ? + NADPH + H+
-
?
(2S,5S)-2,5-hexanediol + NADP+
-
Thermococcus sibiricus DSM 12597 ? + NADPH + H+
-
?
2,3-pentanedione + NADPH + H+
-
Thermococcus sibiricus ? + NADP+
-
?
2,3-pentanedione + NADPH + H+
-
Thermococcus sibiricus DSM 12597 ? + NADP+
-
?
2-propanol + NADP+
-
Thermococcus sibiricus acetone + NADPH + H+
-
?
2-propanol + NADP+
-
Thermococcus sibiricus DSM 12597 acetone + NADPH + H+
-
?
pyruvaldehyde + NADPH + H+
-
Thermococcus sibiricus lactaldehyde + NADP+
-
?
pyruvaldehyde + NADPH + H+
-
Thermococcus sibiricus DSM 12597 lactaldehyde + NADP+
-
?

Subunits

Subunits Comment Organism
tetramer
-
Thermococcus sibiricus

Synonyms

Synonyms Comment Organism
TsAdh319
-
Thermococcus sibiricus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
-
assay at Thermococcus sibiricus
70
-
oxidation of (2S,5S)-2,5-hexanediol, in presence of 1 M NaCl Thermococcus sibiricus
83
-
oxidation of (2S,5S)-2,5-hexanediol, no addition of NaCl Thermococcus sibiricus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
50 75 50°C: about 50% of maximal activity, 75°C: about 60% of maximal activity, oxidation of (2S,5S)-2,5-hexanediol, in presence of 1 M NaCl Thermococcus sibiricus
65 86 65°C: about 50% of maximal activity, 86°C: about 50% of maximal activity, oxidation of (2S,5S)-2,5-hexanediol, no addition of NaCl Thermococcus sibiricus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
thermal denaturation is fully irreversible and follows first-order kinetics Thermococcus sibiricus
80
-
activity begins to decrease after 2 h preincubation. The rate of 2-propanol oxidation is decreased 34% and 58% over 24 h incubation at 80°C in the presence or absence of 1 M NaCl, respectively. Calculated half-life values of the enzyme in buffer with or without 1 M NaCl are 37 vs. 20 h at 80°C, respectively Thermococcus sibiricus
90
-
activity begins to decrease after 30 min preincubation. The incubation at 90°C provokes 31% and 40% reduction of 2-propanol oxidation rate in the presence or absence of 1 M NaCl, respectively. Calculated half-life values of the enzyme in buffer with or without 1 M NaCl are 8 vs. 6 h at 90 °C, respectively Thermococcus sibiricus
102
-
Tm-value in presence of 1 M NaCl Thermococcus sibiricus
104
-
Tm-value in absence of NaCl Thermococcus sibiricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.53
-
2-propanol 60°C, pH 10.5 Thermococcus sibiricus
1.4
-
2,3-Pentanedione 60°C, pH 7.5 Thermococcus sibiricus
1.51
-
NADPH 60°C, pH 7.5, cosubstrate: pyruvaldehyde Thermococcus sibiricus
2.02
-
NADP+ 60°C, pH 10.5, cosubstrate: 2,5-hexandiol Thermococcus sibiricus
2.04
-
Pyruvaldehyde 60°C, pH 7.5 Thermococcus sibiricus
2.2
-
(2S,5S)-2,5-hexanediol 60°C, pH 10.5 Thermococcus sibiricus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Thermococcus sibiricus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0032
-
2-propanol 60°C, pH 10.5 Thermococcus sibiricus
0.11
-
Pyruvaldehyde 60°C, pH 7.5 Thermococcus sibiricus
0.17
-
(2S,5S)-2,5-hexanediol 60°C, pH 10.5 Thermococcus sibiricus
1.33
-
2,3-Pentanedione 60°C, pH 7.5 Thermococcus sibiricus
76
-
NADPH 60°C, pH 7.5, cosubstrate: pyruvaldehyde Thermococcus sibiricus
101
-
NADP+ 60°C, pH 10.5, cosubstrate: 2,5-hexandiol Thermococcus sibiricus