Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Van Zyl, L.J.; Taylor, M.P.; Eley, K.; Tuffin, M.; Cowan, D.A.
    Engineering pyruvate decarboxylase-mediated ethanol production in the thermophilic host Geobacillus thermoglucosidasius (2014), Appl. Microbiol. Biotechnol., 98, 1247-1259.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.1.1 expressed in Escherichia coli BL21(DE3) cells Parageobacillus thermoglucosidasius
4.1.1.1 gene pdc, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha, heterologous expression of the enzyme, passaged through Escherichia coli strain JM109 for DNA methylation, in the thermophile Geobacillus thermoglucosidasius strain TM89, using the the 170-bp promoter region of the lactate dehydrogenase gene Pldh from Geobacillus thermoglucosidasiusNCA1503, for ethanol production Gluconobacter oxydans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.1.1 additional information
-
 additional information the recombinant enzyme expressed in Geobacillus thermoglucosidasius shows normal Michaelis-Menten kinetics with pyruvate Gluconobacter oxydans
4.1.1.1 0.1 2 pyruvate pH 5.0, 55°C, recombinant enzyme Gluconobacter oxydans
4.1.1.1 0.74
-
 pyruvate pH 6.0, 55°C, native enzyme Gluconobacter oxydans
4.1.1.1 1.2
-
 pyruvate pH 6.5, 55°C, recombinant enzyme Gluconobacter oxydans
4.1.1.1 2.8
-
 pyruvate pH 7.0, 55°C, recombinant enzyme Gluconobacter oxydans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.1.1.1 Mg2+ dependent on Parageobacillus thermoglucosidasius

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.1.1.1 59000
-
 x * 60800, about, sequence calculation, x * 59000, recombinant His-tagged enzyme, SDS-PAGEs Gluconobacter oxydans
4.1.1.1 60000
-
 x * 60000, SDS-PAGE Parageobacillus thermoglucosidasius
4.1.1.1 60800
-
-
 Parageobacillus thermoglucosidasius
4.1.1.1 60800
-
 x * 60800, about, sequence calculation, x * 59000, recombinant His-tagged enzyme, SDS-PAGEs Gluconobacter oxydans

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.1.1.1 pyruvate Gluconobacter oxydans
-
 acetaldehyde + CO2
-
 ?
4.1.1.1 pyruvate Parageobacillus thermoglucosidasius
-
 acetaldehyde + CO2
-
 ?
4.1.1.1 pyruvate Parageobacillus thermoglucosidasius TM89
-
 acetaldehyde + CO2
-
 ?
4.1.1.1 pyruvate Gluconobacter oxydans DSM Z 7145
-
 acetaldehyde + CO2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.1.1.1 Gluconobacter oxydans
-
 gene pdc
-
4.1.1.1 Gluconobacter oxydans DSM Z 7145
-
 gene pdc
-
4.1.1.1 Parageobacillus thermoglucosidasius
-
-
-
4.1.1.1 Parageobacillus thermoglucosidasius TM89
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.1.1 HisBind resin column chromatography Parageobacillus thermoglucosidasius
4.1.1.1 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Gluconobacter oxydans

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.1.1.1 4.4
-
 with pyruvate, pH 6.0, 55°C, native enzyme Gluconobacter oxydans
4.1.1.1 47
-
 with pyruvate, pH 6.5, 55°C, recombinant enzyme Gluconobacter oxydans
4.1.1.1 57
-
 with pyruvate, pH 5.0, 55°C, recombinant enzyme Gluconobacter oxydans
4.1.1.1 125
-
 with pyruvate, pH 7.0, 55°C, recombinant enzyme Gluconobacter oxydans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.1.1 2-oxo-4-methylpentanoate
-
 Parageobacillus thermoglucosidasius ?
-
 ?
4.1.1.1 2-oxo-4-methylpentanoate
-
 Parageobacillus thermoglucosidasius TM89 ?
-
 ?
4.1.1.1 2-Oxobutanoate
-
 Parageobacillus thermoglucosidasius ?
-
 ?
4.1.1.1 2-Oxobutanoate
-
 Parageobacillus thermoglucosidasius TM89 ?
-
 ?
4.1.1.1 2-oxobutanoate
-
 Gluconobacter oxydans propionaldehyde + CO2
-
 ?
4.1.1.1 2-oxobutanoate
-
 Gluconobacter oxydans DSM Z 7145 propionaldehyde + CO2
-
 ?
4.1.1.1 2-Oxopentanoate
-
 Parageobacillus thermoglucosidasius ?
-
 ?
4.1.1.1 2-Oxopentanoate
-
 Parageobacillus thermoglucosidasius TM89 ?
-
 ?
4.1.1.1 2-oxopentanoate
-
 Gluconobacter oxydans butyraldehyde + CO2
-
 ?
4.1.1.1 2-oxopentanoate
-
 Gluconobacter oxydans DSM Z 7145 butyraldehyde + CO2
-
 ?
4.1.1.1 4-hydroxyphenylpyruvate
-
 Parageobacillus thermoglucosidasius ?
-
 ?
4.1.1.1 4-methyl-2-oxopentanoate
-
 Gluconobacter oxydans ?
-
 ?
4.1.1.1 4-methyl-2-oxopentanoate
-
 Gluconobacter oxydans DSM Z 7145 ?
-
 ?
4.1.1.1 additional information no activity with 3-phenyl-2-oxopropanoate, benzoyl formate, and indole-3-pyruvate Gluconobacter oxydans ?
-
 ?
4.1.1.1 additional information no activity is recorded on 3-phenyl-2-oxopropanoate, benzoylformate, 4-hydroxyphenylpyruvate, and indole-3-pyruvate Parageobacillus thermoglucosidasius ?
-
 ?
4.1.1.1 additional information no activity is recorded on 3-phenyl-2-oxopropanoate, benzoylformate, 4-hydroxyphenylpyruvate, and indole-3-pyruvate Parageobacillus thermoglucosidasius TM89 ?
-
 ?
4.1.1.1 additional information no activity with 3-phenyl-2-oxopropanoate, benzoyl formate, and indole-3-pyruvate Gluconobacter oxydans DSM Z 7145 ?
-
 ?
4.1.1.1 pyruvate
-
 Gluconobacter oxydans acetaldehyde + CO2
-
 ?
4.1.1.1 pyruvate
-
 Parageobacillus thermoglucosidasius acetaldehyde + CO2
-
 ?
4.1.1.1 pyruvate
-
 Parageobacillus thermoglucosidasius TM89 acetaldehyde + CO2
-
 ?
4.1.1.1 pyruvate
-
 Gluconobacter oxydans DSM Z 7145 acetaldehyde + CO2
-
 ?

Subunits

EC Number Subunits Comment Organism
4.1.1.1 ? x * 60000, SDS-PAGE Parageobacillus thermoglucosidasius
4.1.1.1 ? x * 60800, about, sequence calculation, x * 59000, recombinant His-tagged enzyme, SDS-PAGEs Gluconobacter oxydans
4.1.1.1 ? x * 60800, calculated from amino acid sequence Parageobacillus thermoglucosidasius

Synonyms

EC Number Synonyms Comment Organism
4.1.1.1 PDC
-
 Gluconobacter oxydans
4.1.1.1 PDC
-
 Parageobacillus thermoglucosidasius

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.1.1.1 53
-
-
 Gluconobacter oxydans
4.1.1.1 53
-
 recombinant enzyme Parageobacillus thermoglucosidasius

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
4.1.1.1 25 65 over 50% of maximal activity within this range Gluconobacter oxydans

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.1.1.1 55 80 approximately 40% and 30% enzyme activity is retained after 30 min and 1 h at 65°C, respectively. The native enzyme is stable for 5 min at 55°C but completely inactivated by incubating for 3 min at 80°C Parageobacillus thermoglucosidasius
4.1.1.1 60
-
 at temperatures of or below 60°C the enzyme retains 30% of the initial activity at 65°C after an h of incubation Gluconobacter oxydans
4.1.1.1 65
-
 purified recombinant enzyme, 40% enzyme activity remain after 30 min Gluconobacter oxydans

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.1.1 5 7 pyruvate pH 5.0, 55°C, recombinant enzyme Gluconobacter oxydans
4.1.1.1 47
-
 pyruvate pH 6.5, 55°C, recombinant enzyme Gluconobacter oxydans
4.1.1.1 125
-
 pyruvate pH 7.0, 55°C, recombinant enzyme Gluconobacter oxydans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.1.1.1 4.5 5 recombinant enzyme Gluconobacter oxydans
4.1.1.1 4.5 5 recombinant enzyme Parageobacillus thermoglucosidasius
4.1.1.1 6
-
 native enzyme Gluconobacter oxydans
4.1.1.1 6
-
 native enzyme Parageobacillus thermoglucosidasius

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.1.1 thiamine diphosphate dependent on Parageobacillus thermoglucosidasius

pI Value

EC Number Organism Comment pI Value Maximum pI Value
4.1.1.1 Parageobacillus thermoglucosidasius calculated from amino acid sequence
-
 6
4.1.1.1 Gluconobacter oxydans sequence calculation
-
 6

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.1.1.1 36
-
 pyruvate pH 6.5, 55°C, recombinant enzyme Gluconobacter oxydans
4.1.1.1 42
-
 pyruvate pH 7.0, 55°C, recombinant enzyme Gluconobacter oxydans
4.1.1.1 475
-
 pyruvate pH 5.0, 55°C, recombinant enzyme Gluconobacter oxydans