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Literature summary extracted from

  • Boehme, B.; Moritz, B.; Wendler, J.; Hertel, T.C.; Ihling, C.; Brandt, W.; Pietzsch, M.
    Enzymatic activity and thermoresistance of improved microbial transglutaminase variants (2020), Amino Acids, 52, 313-326 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.2.13 recombinant expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21Gold (DE3) Streptomyces mobaraensis

Protein Variants

EC Number Protein Variants Comment Organism
2.3.2.13 H289Y site-directed mutagenesis Streptomyces mobaraensis
2.3.2.13 K269S site-directed mutagenesis Streptomyces mobaraensis
2.3.2.13 K294L site-directed mutagenesis Streptomyces mobaraensis
2.3.2.13 NG257S site-directed mutagenesis Streptomyces mobaraensis
2.3.2.13 S23Y/S24N site-directed mutagenesis Streptomyces mobaraensis
2.3.2.13 S2P site-directed mutagenesis, the mutant shows increased activity compared to wild-type Streptomyces mobaraensis
2.3.2.13 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.2.13 3.53
-
 CBZ-Gln-Gly-OH recombinant mutant TG16, pH 6.0, 37°C Streptomyces mobaraensis
2.3.2.13 4.02
-
 CBZ-Gln-Gly-OH recombinant mutant S2P, pH 6.0, 37°C Streptomyces mobaraensis
2.3.2.13 8.55
-
 CBZ-Gln-Gly-OH recombinant wild-type enzyme, pH 6.0, 37°C Streptomyces mobaraensis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.2.13 protein glutamine + alkylamine Streptomyces mobaraensis
-
 protein N5-alkylglutamine + NH3
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.2.13 Streptomyces mobaraensis P81453
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.2.13 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21Gold (DE3) by nickel affinity chromatography Streptomyces mobaraensis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.2.13 beta-casein glutamine + hydroxylamine
-
 Streptomyces mobaraensis beta-casein N5-hydroxylglutamine + NH3
-
 ?
2.3.2.13 CBZ-Gln-Gly-OH + hydroxylamine
-
 Streptomyces mobaraensis CBZ-Gln(gamma-monohydroxamate)-Gly + NH3
-
 ?
2.3.2.13 protein glutamine + alkylamine
-
 Streptomyces mobaraensis protein N5-alkylglutamine + NH3
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.2.13 microbial transglutaminase
-
 Streptomyces mobaraensis
2.3.2.13 MTG
-
 Streptomyces mobaraensis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.2.13 37
-
 assay at Streptomyces mobaraensis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.3.2.13 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 [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.2.13 27.37
-
 CBZ-Gln-Gly-OH recombinant wild-type enzyme, pH 6.0, 37°C Streptomyces mobaraensis
2.3.2.13 32.18
-
 CBZ-Gln-Gly-OH recombinant mutant TG16, pH 6.0, 37°C Streptomyces mobaraensis
2.3.2.13 33.92
-
 CBZ-Gln-Gly-OH recombinant mutant S2P, pH 6.0, 37°C Streptomyces mobaraensis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.2.13 6
-
 assay at Streptomyces mobaraensis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.3.2.13 3.2
-
 CBZ-Gln-Gly-OH recombinant wild-type enzyme, pH 6.0, 37°C Streptomyces mobaraensis
2.3.2.13 8.4
-
 CBZ-Gln-Gly-OH recombinant mutant S2P, pH 6.0, 37°C Streptomyces mobaraensis
2.3.2.13 9.1
-
 CBZ-Gln-Gly-OH recombinant mutant TG16, pH 6.0, 37°C Streptomyces mobaraensis