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Literature summary extracted from

  • Sirobhushanam, S.; Galva, C.; Saunders, L.P.; Sen, S.; Jayaswal, R.; Wilkinson, B.J.; Gatto, C.
    Utilization of multiple substrates by butyrate kinase from Listeria monocytogenes (2017), Biochim. Biophys. Acta, 1862, 283-290 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.19 expression in Escherichia coli Listeria monocytogenes
2.7.2.7 expressed in Escherichia coli BL21(DE3) cells Listeria monocytogenes

General Stability

EC Number General Stability Organism
2.3.1.19 loss of catalytic activity due to precipitation of purified Ptb is observed at concentrations ranging from 1-6 mg/ml in 50 mM Tris (pH 7.5) and in 25 mM HEPES (pH 7.5) buffers, which is resolved by storage in high NaCl concentrations (1.3-3 M) Listeria monocytogenes

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.19 0.0315
-
hexanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 0.0315
-
hexanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 0.0438
-
2-ethylbutanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 0.0438
-
2-ethylbutanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 0.0481
-
2-methylbutanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 0.0481
-
2-methylbutanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 0.0532
-
isovaleryl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 0.0532
-
isovaleryl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 0.0579
-
isobutanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 0.0579
-
isobutanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 0.0799
-
butanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 0.0799
-
butanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 0.1608
-
pentanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 0.1608
-
pentanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 0.1906
-
propionyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 0.1906
-
propionyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 0.992
-
acetyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 0.9921
-
acetyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.7.2.7 7.6
-
Isobutyrate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 13.1
-
pentanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 16.5
-
2-ethylbutyrate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 27.3
-
3-methylpentanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 36.2
-
2-methylbutyrate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 44.2
-
2-methylpentanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 50.5
-
Butanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 57.1
-
Isovalerate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 174.1
-
propionate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 1856
-
hexanoate at pH 7.5 and 37°C Listeria monocytogenes

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.2.7 Mn2+ 4 mM used in assay conditions Listeria monocytogenes

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.19 Listeria monocytogenes
-
-
-
2.3.1.19 Listeria monocytogenes 10403S
-
-
-
2.7.2.7 Listeria monocytogenes
-
-
-
2.7.2.7 Listeria monocytogenes 10403S
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.2.7 Ni-NTA resin column chromatography Listeria monocytogenes

Storage Stability

EC Number Storage Stability Organism
2.3.1.19 can be stored for two weeks without detectable loss of activity by dilution of eluted protein to below 0.5 mg/ml in HEPES buffer with 1.5 M NaCl Listeria monocytogenes
2.3.1.19 dilution of eluted protein to below 0.5 mg/ml in HEPES buffer with 1.5 M NaCl, storage for two weeks without detectable loss of activity Listeria monocytogenes

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.19 2-ethylbutanoyl-CoA + phosphate
-
Listeria monocytogenes 2-ethylbutanoyl phosphate + CoA
-
?
2.3.1.19 2-ethylbutanoyl-CoA + phosphate
-
Listeria monocytogenes CoA + 2-ethylbutanoyl phosphate
-
?
2.3.1.19 2-methylbutanoyl-CoA + phosphate
-
Listeria monocytogenes 2-methylbutanoyl phosphate + CoA
-
?
2.3.1.19 2-methylbutanoyl-CoA + phosphate
-
Listeria monocytogenes CoA + 2-methylbutanoyl phosphate
-
?
2.3.1.19 acetyl-CoA + phosphate
-
Listeria monocytogenes CoA + acetyl phosphate
-
?
2.3.1.19 acetyl-CoA + phosphate
-
Listeria monocytogenes 10403S CoA + acetyl phosphate
-
?
2.3.1.19 acetyl-CoA + phosphate
-
Listeria monocytogenes acetyl Phosphate + CoA
-
?
2.3.1.19 acetyl-CoA + phosphate
-
Listeria monocytogenes 10403S acetyl Phosphate + CoA
-
?
2.3.1.19 butanoyl-CoA + phosphate
-
Listeria monocytogenes butanoyl phosphate + CoA
-
?
2.3.1.19 butanoyl-CoA + phosphate
-
Listeria monocytogenes CoA + butanoyl phosphate
-
?
2.3.1.19 hexanoyl-CoA + phosphate
-
Listeria monocytogenes CoA + hexanoyl phosphate
-
?
2.3.1.19 hexanoyl-CoA + phosphate
-
Listeria monocytogenes hexanoyl phosphate + CoA
-
?
2.3.1.19 isobutanoyl-CoA + phosphate
-
Listeria monocytogenes CoA + isobutanoyl phosphate
-
?
2.3.1.19 isobutanoyl-CoA + phosphate
-
Listeria monocytogenes isobutanoyl phosphate + CoA
-
?
2.3.1.19 isovaleryl-CoA + phosphate
-
Listeria monocytogenes CoA + isovaleryl phosphate
-
?
2.3.1.19 isovaleryl-CoA + phosphate
-
Listeria monocytogenes isovaleryl phosphate + CoA
-
?
2.3.1.19 additional information enzyme exhibits a broad substrate specificity, a strong preference for branched-chain substrates, a lack of activity with acetyl CoA and hexanoyl CoA, and strict chain length preference (C3-C5). Catalysis involves ternary complex formation Listeria monocytogenes ?
-
-
2.3.1.19 additional information mechanism is consistent with ternary complex formation rather than a ping-pong mechanism. PtB is active with a large number of substrates and prefers substrates with a carbon chain length of C3-C5 and the presence of an alkyl side chain Listeria monocytogenes ?
-
-
2.3.1.19 additional information enzyme exhibits a broad substrate specificity, a strong preference for branched-chain substrates, a lack of activity with acetyl CoA and hexanoyl CoA, and strict chain length preference (C3-C5). Catalysis involves ternary complex formation Listeria monocytogenes 10403S ?
-
-
2.3.1.19 additional information mechanism is consistent with ternary complex formation rather than a ping-pong mechanism. PtB is active with a large number of substrates and prefers substrates with a carbon chain length of C3-C5 and the presence of an alkyl side chain Listeria monocytogenes 10403S ?
-
-
2.3.1.19 pentanoyl-CoA + phosphate
-
Listeria monocytogenes CoA + pentanoyl phosphate
-
?
2.3.1.19 pentanoyl-CoA + phosphate
-
Listeria monocytogenes pentanoyl phosphate + CoA
-
?
2.3.1.19 propionyl-CoA + phosphate
-
Listeria monocytogenes CoA + propionyl phosphate
-
?
2.3.1.19 propionyl-CoA + phosphate
-
Listeria monocytogenes 10403S CoA + propionyl phosphate
-
?
2.3.1.19 propionyl-CoA + phosphate
-
Listeria monocytogenes propionyl phosphate + CoA
-
?
2.7.2.7 ATP + 2-ethylbutyrate
-
Listeria monocytogenes ADP + 2-ethylbutyryl phosphate
-
?
2.7.2.7 ATP + 2-methylbutyrate
-
Listeria monocytogenes ADP + 2-methylbutyryl phosphate
-
?
2.7.2.7 ATP + 2-methylpentanoate
-
Listeria monocytogenes ADP + 2-methylpentanoyl phosphate
-
?
2.7.2.7 ATP + 3-methylpentanoate
-
Listeria monocytogenes ADP + 3-methylpentanoyl phosphate
-
?
2.7.2.7 ATP + acetate
-
Listeria monocytogenes ADP + acetyl phosphate
-
?
2.7.2.7 ATP + butanoate third best substrate Listeria monocytogenes ADP + butanoyl phosphate
-
?
2.7.2.7 ATP + hexanoate
-
Listeria monocytogenes ADP + hexanoyl phosphate
-
?
2.7.2.7 ATP + isobutyrate second best substrate Listeria monocytogenes ADP + isobutyroyl phosphate
-
?
2.7.2.7 ATP + isovalerate
-
Listeria monocytogenes ADP + isovaleroyl phosphate
-
?
2.7.2.7 ATP + pentanoate best substrate Listeria monocytogenes ADP + pentanoyl phosphate
-
?
2.7.2.7 ATP + propionate
-
Listeria monocytogenes ADP + propionyl phosphate
-
?

Subunits

EC Number Subunits Comment Organism
2.3.1.19 ? x * 34000, SDS-PAGE Listeria monocytogenes
2.7.2.7 ? x * 42000, SDS-PAGE Listeria monocytogenes

Synonyms

EC Number Synonyms Comment Organism
2.3.1.19 PTB
-
Listeria monocytogenes
2.7.2.7 Buk
-
Listeria monocytogenes

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.1.19 47
-
hexanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 47
-
hexanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 100.9
-
2-ethylbutanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 100.9
-
2-ethylbutanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 302.4
-
acetyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 302.4
-
acetyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 518.8
-
propionyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 518.8
-
propionyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 673.9
-
butanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 673.9
-
butanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 676.1
-
pentanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 676.1
-
pentanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 849.1
-
2-methylbutanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 849.1
-
2-methylbutanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 1043.4
-
isovaleryl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 1043.4
-
isovaleryl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 1263.4
-
isobutanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 1263.4
-
isobutanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.7.2.7 0.11
-
2-ethylbutyrate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.27
-
2-methylbutyrate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.31
-
2-methylpentanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.35
-
3-methylpentanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.4
-
Isobutyrate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.42
-
Isovalerate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.84
-
Butanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 1.2
-
pentanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 2.59
-
propionate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 21.85
-
hexanoate at pH 7.5 and 37°C Listeria monocytogenes

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.3.1.19 300
-
acetyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 300
-
acetyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 1700
-
hexanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 1700
-
hexanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 2300
-
2-ethylbutanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 2720
-
propionyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 2720
-
propionyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 4200
-
pentanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 4200
-
pentanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 8430
-
butanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 8430
-
butanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 17700
-
2-methylbutanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 17700
-
2-methylbutanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 19600
-
isovaleryl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 19600
-
isovaleryl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 21400
-
isobutanoyl-CoA pH 7.5, 25°C Listeria monocytogenes
2.3.1.19 21400
-
isobutanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.3.1.19 23000
-
2-ethylbutanoyl-CoA pH 7.5, 23°C Listeria monocytogenes
2.7.2.7 0.007
-
Isovalerate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.007
-
2-methylpentanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.007
-
2-ethylbutyrate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.011
-
hexanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.013
-
3-methylpentanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.015
-
propionate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.017
-
Butanoate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.052
-
Isobutyrate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.07
-
2-methylbutyrate at pH 7.5 and 37°C Listeria monocytogenes
2.7.2.7 0.092
-
pentanoate at pH 7.5 and 37°C Listeria monocytogenes