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Literature summary for 1.1.1.6 extracted from

  • Zhang, H.; Lountos, G.T.; Ching, C.B.; Jiang, R.
    Engineering of glycerol dehydrogenase for improved activity towards 1, 3-butanediol (2010), Appl. Microbiol. Biotechnol., 88, 117-124.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
His-tagged version expressed in Escherichia coli BL21(DE3) Escherichia coli
His-tagged version expressed in Escherichia coli BL21(DE3) Klebsiella pneumoniae
His-tagged version expressed in Escherichia coli BL21(DE3) Salmonella enterica

Protein Variants

Protein Variants Comment Organism
A15T/D16G/V17A/N19K/E23D/Q45E/S46E/T47M/V48L/E49R/F52L/K53A/V58A/V59A/Q70H/D74N/G78D/E81G/T82N/Q83K/G86T/I88V/G108N/R139S/L142M/N145R/K155Q/V256I/L260M mutant selected from a DNA shuffling library (Escherichia coli, Salmonella enterica, Klebsiella pneumoniae) Escherichia coli
D121A D121 can potentially hinder the proper binding of substrate 1,3-butanediol due to steric hindrance Escherichia coli
D16N/N19A/E23D/L28M/E30N/R31N/Q45E/S46E/V48L/E49R/F52L/K53T/D54G/V58S/G78V/I79V/T82K/A83S/I88V/G108N/R139S/L142M/N145R/K155Q/L211I/G248S/V256I/H268Y/D317E/P319L mutant selected from a DNA shuffling library (Escherichia coli, Salmonella enterica, Klebsiella pneumoniae) Escherichia coli
Q70H/D121A/G193C/E291Q/A310T mutant selected from a DNA shuffling library (Escherichia coli, Salmonella enterica, Klebsiella pneumoniae) and site-directed mutation D121A Escherichia coli
Q70H/D74N/G78D/E81G/T82N/Q83K/C84Y/G86T/I88V/G108N/E134A/E204K/L211I/E215K/I234V/V256I/L260M/E291D/S300C/A302S/E316G/V318I/A320T/I324L/T344D/P345S mutant selected from a DNA shuffling library (Escherichia coli, Salmonella enterica, Klebsiella pneumoniae) Escherichia coli
Q70H/G193C/E291Q/A310T mutant selected from a DNA shuffling library (Escherichia coli, Salmonella enterica, Klebsiella pneumoniae) Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0165
-
NAD+ wild type protein, pH 9.5, 30°C Escherichia coli
0.0182
-
NAD+ mutant protein Q70H/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli
0.4614
-
NAD+ mutant protein Q70H/D121A/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli
0.5946
-
NAD+ mutant protein D121A, pH 9.5, 30°C Escherichia coli
11.9
-
1,3-butanediol mutant protein Q70H/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli
16.2
-
1,3-butanediol wild type protein, pH 9.5, 30°C Escherichia coli
20.9
-
1,3-butanediol pH 9.5, 30°C Klebsiella pneumoniae
29.2
-
1,3-butanediol pH 9.5, 30°C Salmonella enterica
49.9
-
1,3-butanediol mutant protein D121A, pH 9.5, 30°C Escherichia coli
77.8
-
1,3-butanediol mutant protein Q70H/D121A/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+
-
Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
39000
-
x * 39000, SDS-PAGE Escherichia coli
39000
-
x * 39000, SDS-PAGE Klebsiella pneumoniae
39000
-
x * 39000, SDS-PAGE Salmonella enterica

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-
Klebsiella pneumoniae
-
-
-
Klebsiella pneumoniae MGH78578
-
-
-
Salmonella enterica
-
-
-
Salmonella enterica ATCC 9150
-
-
-

Purification (Commentary)

Purification (Comment) Organism
immobilized metal ion affinity chromatography (Co2+) Escherichia coli
immobilized metal ion affinity chromatography (Co2+) Klebsiella pneumoniae
immobilized metal ion affinity chromatography (Co2+) Salmonella enterica

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.7
-
pH 9.5, 30°C Klebsiella pneumoniae
3.9
-
pH 9.5, 30°C Salmonella enterica
4.8
-
wild type protein, pH 9.5, 30°C Escherichia coli
6
-
mutant protein D16N/N19A/E23D/L28M/E30N/R31N/Q45E/S46E/V48L/E49R/F52L/K53T/D54G/V58S/G78V/I79V/T82K/A83S/I88V/G108N/R139S/L142M/N145R/K155Q/L211I/G248S/V256I/H268Y/D317E/P319L, pH 9.5, 30°C Escherichia coli
6.8
-
mutant protein A15T/D16G/V17A/N19K/E23D/Q45E/S46E/T47M/V48L/E49R/F52L/K53A/V58A/V59A/Q70H/D74N/G78D/E81G/T82N/Q83K/G86T/I88V/G108N/R139S/L142M/N145R/K155Q/V256I/L260M, pH 9.5, 30°C Escherichia coli
7.4
-
mutant protein Q70H/D74N/G78D/E81G/T82N/Q83K/C84Y/G86T/I88V/G108N/E134A/E204K/L211I/E215K/I234V/V256I/L260M/E291D/S300C/A302S/E316G/V318I/A320T/I324L/T344D/P345S, pH 9.5, 30°C Escherichia coli
13.2
-
mutant protein Q70H/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli
90.7
-
mutant protein D121A, pH 9.5, 30°C Escherichia coli
126.6
-
mutant protein Q70H/D121A/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1,3-butanediol + NAD+
-
Escherichia coli 4-hydroxy-2-butanone + NADH + H+
-
?
1,3-butanediol + NAD+
-
Klebsiella pneumoniae 4-hydroxy-2-butanone + NADH + H+
-
?
1,3-butanediol + NAD+
-
Salmonella enterica 4-hydroxy-2-butanone + NADH + H+
-
?
1,3-butanediol + NAD+
-
Salmonella enterica ATCC 9150 4-hydroxy-2-butanone + NADH + H+
-
?
1,3-butanediol + NAD+
-
Klebsiella pneumoniae MGH78578 4-hydroxy-2-butanone + NADH + H+
-
?

Subunits

Subunits Comment Organism
? x * 39000, SDS-PAGE Escherichia coli
? x * 39000, SDS-PAGE Klebsiella pneumoniae
? x * 39000, SDS-PAGE Salmonella enterica

Synonyms

Synonyms Comment Organism
glycerol dehydrogenase
-
Escherichia coli
glycerol dehydrogenase
-
Klebsiella pneumoniae
glycerol dehydrogenase
-
Salmonella enterica

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.6
-
1,3-butanediol pH 9.5, 30°C Klebsiella pneumoniae
2.7
-
1,3-butanediol pH 9.5, 30°C Salmonella enterica
3.3
-
1,3-butanediol wild type protein, pH 9.5, 30°C Escherichia coli
9.1
-
1,3-butanediol mutant protein Q70H/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli
62.4
-
1,3-butanediol mutant protein D121A, pH 9.5, 30°C Escherichia coli
87.2
-
1,3-butanediol mutant protein Q70H/D121A/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.09
-
1,3-butanediol pH 9.5, 30°C Salmonella enterica
0.12
-
1,3-butanediol pH 9.5, 30°C Klebsiella pneumoniae
0.2
-
1,3-butanediol wild type protein, pH 9.5, 30°C Escherichia coli
0.77
-
1,3-butanediol mutant protein Q70H/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli
1.12
-
1,3-butanediol mutant protein Q70H/D121A/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli
1.26
-
1,3-butanediol mutant protein D121A, pH 9.5, 30°C Escherichia coli
105
-
NAD+ mutant protein D121A, pH 9.5, 30°C Escherichia coli
188
-
NAD+ mutant protein Q70H/D121A/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli
200
-
NAD+ wild type protein, pH 9.5, 30°C Escherichia coli
500
-
NAD+ mutant protein Q70H/G193C/E291Q/A310T, pH 9.5, 30°C Escherichia coli