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Literature summary for 4.1.1.1 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
Search for more literature for 4.1.1.1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Parageobacillus thermoglucosidasius
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]

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

Metals/Ions

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

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

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms Comment Organism
PDC
-
 Gluconobacter oxydans
PDC
-
 Parageobacillus thermoglucosidasius

Temperature Optimum [°C]

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

Temperature Range [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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
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
65
-
 purified recombinant enzyme, 40% enzyme activity remain after 30 min Gluconobacter oxydans

Turnover Number [1/s]

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

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
thiamine diphosphate dependent on Parageobacillus thermoglucosidasius

pI Value

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

kcat/KM [mM/s]

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