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

BRENDA support

Literature summary for 2.3.2.13 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)

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

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 [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/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Streptomyces mobaraensis P81453
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms Comment Organism
microbial transglutaminase
-
Streptomyces mobaraensis
MTG
-
Streptomyces mobaraensis

Temperature Optimum [°C]

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

Temperature Stability [°C]

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 [1/s]

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

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

kcat/KM [mM/s]

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