Cloned (Comment) | Organism |
---|---|
recombinant expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21Gold (DE3) | Streptomyces mobaraensis |
Protein Variants | Comment | Organism |
---|---|---|
H289Y | site-directed mutagenesis | Streptomyces mobaraensis |
K269S | site-directed mutagenesis | Streptomyces mobaraensis |
K294L | site-directed mutagenesis | Streptomyces mobaraensis |
NG257S | site-directed mutagenesis | Streptomyces mobaraensis |
S23Y/S24N | site-directed mutagenesis | Streptomyces mobaraensis |
S2P | site-directed mutagenesis, the mutant shows increased activity compared to wild-type | Streptomyces mobaraensis |
S2P/S23Y/S24N/H289Y/K294L | site-directed mutagenesis, the mutant TG16 shows 19fold reduced thermal stability/half-life at 60°C compared to wild-type enzyme, differential scanning fluorimetry, the transition point of thermal unfolding is increased by 7.9°C compared to wild-type. The inactivation process follows a pseudo-first-order reaction which is accompanied by irreversible aggregation and intramolecular self-crosslinking of the enzyme. The increased thermoresistance is caused by a higher backbone rigidity as well as increased hydrophobic interactions and newly formed hydrogen bridges, molecular dynamics simulations, overview. The mutant shows increased activity compared to wild-type | Streptomyces mobaraensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.53 | - |
CBZ-Gln-Gly-OH | recombinant mutant TG16, pH 6.0, 37°C | Streptomyces mobaraensis | |
4.02 | - |
CBZ-Gln-Gly-OH | recombinant mutant S2P, pH 6.0, 37°C | Streptomyces mobaraensis | |
8.55 | - |
CBZ-Gln-Gly-OH | recombinant wild-type enzyme, pH 6.0, 37°C | Streptomyces mobaraensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
protein glutamine + alkylamine | Streptomyces mobaraensis | - |
protein N5-alkylglutamine + NH3 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces mobaraensis | P81453 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21Gold (DE3) by nickel affinity chromatography | Streptomyces mobaraensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
beta-casein glutamine + hydroxylamine | - |
Streptomyces mobaraensis | beta-casein N5-hydroxylglutamine + NH3 | - |
? | |
CBZ-Gln-Gly-OH + hydroxylamine | - |
Streptomyces mobaraensis | CBZ-Gln(gamma-monohydroxamate)-Gly + NH3 | - |
? | |
protein glutamine + alkylamine | - |
Streptomyces mobaraensis | protein N5-alkylglutamine + NH3 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
microbial transglutaminase | - |
Streptomyces mobaraensis |
MTG | - |
Streptomyces mobaraensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Streptomyces mobaraensis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
the S2P/S23Y/S24N/H289Y/K294L mutant shows 19fold reduced thermal stability/half-life at 60°C compared to wild-type enzyme, differential scanning fluorimetry, the transition point of thermal unfolding is increased by 7.9°C compared to wild-type. The inactivation process follows a pseudo-first-order reaction which is accompanied by irreversible aggregation and intramolecular self-crosslinking of the enzyme. The increased thermoresistance is caused by a higher backbone rigidity as well as increased hydrophobic interactions and newly formed hydrogen bridges, molecular dynamics simulations, kinetics, overview | Streptomyces mobaraensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
27.37 | - |
CBZ-Gln-Gly-OH | recombinant wild-type enzyme, pH 6.0, 37°C | Streptomyces mobaraensis | |
32.18 | - |
CBZ-Gln-Gly-OH | recombinant mutant TG16, pH 6.0, 37°C | Streptomyces mobaraensis | |
33.92 | - |
CBZ-Gln-Gly-OH | recombinant mutant S2P, pH 6.0, 37°C | Streptomyces mobaraensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Streptomyces mobaraensis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.2 | - |
CBZ-Gln-Gly-OH | recombinant wild-type enzyme, pH 6.0, 37°C | Streptomyces mobaraensis | |
8.4 | - |
CBZ-Gln-Gly-OH | recombinant mutant S2P, pH 6.0, 37°C | Streptomyces mobaraensis | |
9.1 | - |
CBZ-Gln-Gly-OH | recombinant mutant TG16, pH 6.0, 37°C | Streptomyces mobaraensis |