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Literature summary for 2.7.7.72 extracted from

  • Ernst, F.G.M.; Erber, L.; Sammler, J.; Juehling, F.; Betat, H.; Moerl, M.
    Cold adaptation of tRNA nucleotidyltransferases A tradeoff in activity, stability and fidelity (2018), RNA Biol., 15, 144-155 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.7.72

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Bacillus subtilis
expression in Escherichia coli Exiguobacterium sibiricum
expression in Escherichia coli Geobacillus stearothermophilus
expression in Escherichia coli Planococcus halocryophilus

Protein Variants

Protein Variants Comment Organism
additional information exchange of the N- and C-termini (carrying the catalytic core and the tRNA binding region, respectively) of Exiguobacterium sibiricum and Geobacillus stearothermophilus enzymes. The chimera show a reaction temperature optimum comparable to that of the parental cold-adapted enzyme, and both show a single-step denaturation curve with melting temperatures of 55 to 57°C Exiguobacterium sibiricum
additional information exchange of the N- and C-termini (carrying the catalytic core and the tRNA binding region, respectively) of Exiguobacterium sibiricum and Geobacillus stearothermophilus enzymes. The chimera show a reaction temperature optimum comparable to that of the parental cold-adapted enzyme, and both show a single-step denaturation curve with melting temperatures of 55 to 57°C Geobacillus stearothermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.004
-
 ATP pH 7.6, 25°C Geobacillus stearothermophilus
0.005
-
 ATP pH 7.6, 25°C Bacillus subtilis
0.009
-
 ATP pH 7.6, 4°C Bacillus subtilis
0.012
-
 ATP pH 7.6, 40°C Bacillus subtilis
0.013
-
 ATP pH 7.6, 4°C Geobacillus stearothermophilus
0.02
-
 ATP pH 7.6, 55°C Geobacillus stearothermophilus
0.059
-
 ATP pH 7.6, 25°C Exiguobacterium sibiricum
0.072
-
 ATP pH 7.6, 25°C Planococcus halocryophilus
0.843
-
 ATP pH 7.6, 4°C Planococcus halocryophilus
0.934
-
 ATP pH 7.6, 4°C Exiguobacterium sibiricum

Organism

Organism UniProt Comment Textmining
Bacillus subtilis P42977
-
-
Bacillus subtilis 168 P42977
-
-
Exiguobacterium sibiricum B1YHR1
-
-
Exiguobacterium sibiricum DSM 17290 B1YHR1
-
-
Geobacillus stearothermophilus Q7SIB1
-
-
Planococcus halocryophilus A0A1C7DQ98
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
tRNA(Phe) precursor + 2 CTP + ATP substrate is tRNA(Phe) precursor from Saccharomyces cerevisiae Bacillus subtilis tRNA(Phe) with a 3' CCA end + 3 diphosphate
-
 ?
tRNA(Phe) precursor + 2 CTP + ATP substrate is tRNA(Phe) precursor from Saccharomyces cerevisiae Exiguobacterium sibiricum tRNA(Phe) with a 3' CCA end + 3 diphosphate
-
 ?
tRNA(Phe) precursor + 2 CTP + ATP substrate is tRNA(Phe) precursor from Saccharomyces cerevisiae Geobacillus stearothermophilus tRNA(Phe) with a 3' CCA end + 3 diphosphate
-
 ?
tRNA(Phe) precursor + 2 CTP + ATP substrate is tRNA(Phe) precursor from Saccharomyces cerevisiae Planococcus halocryophilus tRNA(Phe) with a 3' CCA end + 3 diphosphate
-
 ?
tRNA(Phe) precursor + 2 CTP + ATP substrate is tRNA(Phe) precursor from Saccharomyces cerevisiae Bacillus subtilis 168 tRNA(Phe) with a 3' CCA end + 3 diphosphate
-
 ?
tRNA(Phe) precursor + 2 CTP + ATP substrate is tRNA(Phe) precursor from Saccharomyces cerevisiae Exiguobacterium sibiricum DSM 17290 tRNA(Phe) with a 3' CCA end + 3 diphosphate
-
 ?

Synonyms

Synonyms Comment Organism
BBI08_05760
-
 Planococcus halocryophilus
Exig_1782
-
 Exiguobacterium sibiricum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
20 30
-
 Bacillus subtilis
20 30
-
 Exiguobacterium sibiricum
20 30
-
 Planococcus halocryophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
33
-
 melting temperature Exiguobacterium sibiricum
38
-
 melting temperature Bacillus subtilis
38
-
 melting temperature Planococcus halocryophilus
73
-
 melting temperature Geobacillus stearothermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0057
-
 ATP pH 7.6, 4°C Geobacillus stearothermophilus
0.036
-
 ATP pH 7.6, 4°C Bacillus subtilis
0.08
-
 ATP pH 7.6, 4°C Exiguobacterium sibiricum
0.092
-
 ATP pH 7.6, 25°C Geobacillus stearothermophilus
0.103
-
 ATP pH 7.6, 4°C Planococcus halocryophilus
0.14
-
 ATP pH 7.6, 25°C Planococcus halocryophilus
0.149
-
 ATP pH 7.6, 25°C Exiguobacterium sibiricum
0.201
-
 ATP pH 7.6, 25°C Bacillus subtilis
0.69
-
 ATP pH 7.6, 40°C Bacillus subtilis
1.58
-
 ATP pH 7.6, 55°C Geobacillus stearothermophilus

General Information

General Information Comment Organism
metabolism comparison of the corresponding enzymes of Exiguobacterium sibiricum that is able to tolerate temperatures of -2.5°C and Planococcus halocryophilus, growing at -15°C, and those of Bacillus subtilis subsp. subtilis and Geobacillus stearothermophilus Bacillus subtilis
metabolism comparison of the corresponding enzymes of Exiguobacterium sibiricum that is able to tolerate temperatures of -2.5°C and Planococcus halocryophilus, growing at -15°C, and those of Bacillus subtilis subsp. subtilis and Geobacillus stearothermophilus. The cold-adapted polymerases show a remarkable error rate during CCA synthesis in vitro as well as in vivo. CCA-adding activity at low temperatures is achieved at the expense of structural stability, and results in a reduced polymerization fidelity Exiguobacterium sibiricum
metabolism comparison of the corresponding enzymes of Exiguobacterium sibiricum that is able to tolerate temperatures of -2.5°C and Planococcus halocryophilus, growing at -15°C, and those of Bacillus subtilis subsp. subtilis and Geobacillus stearothermophilus. The cold-adapted polymerases show a remarkable error rate during CCA synthesis in vitro as well as in vivo. CCA-adding activity at low temperatures is achieved at the expense of structural stability, and results in a reduced polymerization fidelity Geobacillus stearothermophilus
metabolism comparison of the corresponding enzymes of Exiguobacterium sibiricum that is able to tolerate temperatures of -2.5°C and Planococcus halocryophilus, growing at -15°C, and those of Bacillus subtilis subsp. subtilis and Geobacillus stearothermophilus. The cold-adapted polymerases show a remarkable error rate during CCA synthesis in vitro as well as in vivo. CCA-adding activity at low temperatures is achieved at the expense of structural stability, and results in a reduced polymerization fidelity Planococcus halocryophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.086
-
 ATP pH 7.6, 4°C Exiguobacterium sibiricum
0.122
-
 ATP pH 7.6, 4°C Planococcus halocryophilus
0.432
-
 ATP pH 7.6, 4°C Geobacillus stearothermophilus
1.944
-
 ATP pH 7.6, 25°C Planococcus halocryophilus
2.614
-
 ATP pH 7.6, 25°C Exiguobacterium sibiricum
4.138
-
 ATP pH 7.6, 4°C Bacillus subtilis
26.286
-
 ATP pH 7.6, 25°C Geobacillus stearothermophilus
37.925
-
 ATP pH 7.6, 25°C Bacillus subtilis
57.5
-
 ATP pH 7.6, 40°C Bacillus subtilis
79
-
 ATP pH 7.6, 55°C Geobacillus stearothermophilus