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Literature summary extracted from

  • Javitt, G.; Ben-Barak-Zelas, Z.; Jerabek-Willemsen, M.; Fishman, A.
    Constitutive expression of active microbial transglutaminase in Escherichia coli and comparative characterization to a known variant (2017), BMC Biotechnol., 17, 23 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.2.13 functional recombinant expression of wild-type and engineered enzyme in Escherichia coli strain BL21(DE3), improvement of cloning and constitutive expression of soluble active mTG, overview. Usage of constitutive vector pET9a and a synthetic construct encoding the C-terminally His-tagged mTG thermostable variant S2P. Further increase in expression levels may be possible by coexpressing periplasmic secretory proteins, since secretion into the periplasm is frequently the limiting factor for production Streptomyces mobaraensis

Protein Variants

EC Number Protein Variants Comment Organism
2.3.2.13 additional information the S2P variant is generated by random mutagenesis of the wild-type enzyme, and found to be more thermostable, able to withstand incubation at 60°C, and more active than the wild-type enzyme. The synthetic operon construct (based on GenBank ID KX775947) consists of two parts: first a gene encoding the pro-domain crucial for proper folding of the enzyme and second the gene encoding the mTG thermostable variant S2P, with a C-terminal His-tag. Each part is paired with a preceding PelB secretory sequence. The Km value is 3fold lower for the mutant S2P as compared to the wild-type. Conversely, the turnover number is higher for the wild-type enzyme, although the enzymatic efficiency is 2fold higher for the mutant. The mutant unfolds at a slightly higher temperature (56.3°C vs. 55.8°C) indicating improved thermostability although not statistically significant Streptomyces mobaraensis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.2.13 additional information
-
 additional information Michaelis-Menten kinetics. The Km value is 3fold lower for the mutant S2P as compared to the wild-type. Conversely, the turnover number is higher for the wild-type enzyme, although the enzymatic efficiency is 2fold higher for the mutant Streptomyces mobaraensis
2.3.2.13 4.2
-
 Z-Gln-Gly recombinant mutant S2P, pH 7.2, 37°C Streptomyces mobaraensis
2.3.2.13 11.6
-
 Z-Gln-Gly recombinant wild-type enzyme, pH 7.2, 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 additional information Streptomyces mobaraensis transglutaminase catalyzes the acyl transfer reaction between gamma-carboxyamide groups (acyl donor) and primary amines (acyl acceptor). In proteins, it is able to crosslink the gamma-carboxyamide of glutamine and the primary epsilon-amine in lysine ?
-
-
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 BL21(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 additional information transglutaminase catalyzes the acyl transfer reaction between gamma-carboxyamide groups (acyl donor) and primary amines (acyl acceptor). In proteins, it is able to crosslink the gamma-carboxyamide of glutamine and the primary epsilon-amine in lysine Streptomyces mobaraensis ?
-
-
2.3.2.13 additional information the enzymatic transamidation reaction between a gamma-glutamyl donor (Z-Gln-Gly) and hydroxylamine releasing ammonia is coupled to the glutamate dehydrogenase (GDH)-catalyzed reductive amination of 2-oxoglutarate. The activity of GDH is dependent on NADH as a cofactor, whose disappearance can be monitored at 340 nm. NADH concentration was calculated based on a calibration curve, which in turn is used to calculate activity of mTG Streptomyces mobaraensis ?
-
-
2.3.2.13 protein glutamine + alkylamine
-
 Streptomyces mobaraensis protein N5-alkylglutamine + NH3
-
 ?
2.3.2.13 Z-Gln-Gly + hydroxylamine
-
 Streptomyces mobaraensis Z-N5-hydroxyglutaminyl-Gly + NH3
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.2.13 microbial transglutaminase
-
 Streptomyces mobaraensis
2.3.2.13 MTG
-
 Streptomyces mobaraensis
2.3.2.13 TGase
-
 Streptomyces mobaraensis
2.3.2.13 transglutaminase
-
 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 45.5 56.3 melting temperature of enzyme mutant S2P Streptomyces mobaraensis
2.3.2.13 55.8
-
 melting temperature of wild-type enzyme 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 0.6
-
 Z-Gln-Gly recombinant mutant S2P, pH 7.2, 37°C Streptomyces mobaraensis
2.3.2.13 0.85
-
 Z-Gln-Gly recombinant wild-type enzyme, pH 7.2, 37°C Streptomyces mobaraensis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.2.13 7.2
-
 assay at Streptomyces mobaraensis

General Information

EC Number General Information Comment Organism
2.3.2.13 physiological function microbial transglutaminase (mTG) is a robust enzyme catalyzing the formation of an isopeptide bond between glutamine and lysine residues. Transglutaminase catalyzes the acyl transfer reaction between gamma-carboxyamide groups (acyl donor) and primary amines (acyl acceptor). In proteins, it is able to crosslink the gamma-carboxyamide of glutamine and the primary epsilon-amine in lysine 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 0.073
-
 Z-Gln-Gly recombinant wild-type enzyme, pH 7.2, 37°C Streptomyces mobaraensis
2.3.2.13 0.143
-
 Z-Gln-Gly recombinant mutant S2P, pH 7.2, 37°C Streptomyces mobaraensis