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Literature summary for 2.3.2.13 extracted from

  • Wan, W.; He, D.; Xue, Z.; Zhang, Z.
    Specific mutation of transglutaminase gene from Streptomyces hygroscopicus H197 and characterization of microbial transglutaminase (2017), J. Biosci., 42, 537-546 .
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information the inactive proform of the enzyme is activated by cleavage through trypsin Streptomyces hygroscopicus

Cloned(Commentary)

Cloned (Comment) Organism
gene mtg, cloning and recombinant expression of His-tagged wild-type and truncated mutant enzymes in pro-form and inactive zymogen in Escherichia coli strain Rosetta (DE3) Streptomyces hygroscopicus

Protein Variants

Protein Variants Comment Organism
additional information the enzyme is mutated by deleting a specific 84 bp fragment using overlapping extension PCR. The deletion of 28 amino acid residues fragment presents an external free state that results in the mutant MTG spatial configuration to be compressed and thus can enhance the stability or solubility of mutant MTG. The mutant MTG is more stable than the wild-type MTG at 50-60°C, at pH 4.0-9.0, at 7-9% salinity, 30-35% ethanol concentration, and in the presence of Li(I) and Ag(I). The mutant MTG is an intracellular expression protein and mainly expresses in soluble form Streptomyces hygroscopicus

General Stability

General Stability Organism
wild-type and truncated mutant MTGs show higher stability at 40°C and 1-3% salinity, polyhydroxy compounds, i.e. sucrose, cellulose, and PEG, as protein stabilizers can elevate MTG stability Streptomyces hygroscopicus

Inhibitors

Inhibitors Comment Organism Structure
EDTA
-
Streptomyces hygroscopicus
additional information the amount of ammonia produced by cross-linking casein with the mutant MTG in the presence of 20% ethanol is less than the control Streptomyces hygroscopicus

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular the enzyme is secreted Streptomyces hygroscopicus
-
-
additional information the truncated mutant enzyme remains an intracellular protein Streptomyces hygroscopicus
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ required Streptomyces hygroscopicus
NaCl wild-type and truncated mutant MTGs show higher stability at 1-3% salinity Streptomyces hygroscopicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
protein glutamine + alkylamine Streptomyces hygroscopicus
-
protein N5-alkylglutamine + NH3
-
?
protein glutamine + alkylamine Streptomyces hygroscopicus H197
-
protein N5-alkylglutamine + NH3
-
?

Organic Solvent Stability

Organic Solvent Comment Organism
Ethanol wild-type and truncated mutant MTGs are strongly affected by ethanol. The presence of 20% ethanol does not evidently influence the cross-linking reaction of casein catalysed by the mutant MTG Streptomyces hygroscopicus

Organism

Organism UniProt Comment Textmining
Streptomyces hygroscopicus A0A291LTQ0
-
-
Streptomyces hygroscopicus H197 A0A291LTQ0
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification the inactive proform of the enzyme is activated by cleavage through trypsin Streptomyces hygroscopicus

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and truncated mutant enzymes from Escherichia coli by nickel affinity chromatography, activation of inactive pro-MTG (mutant MTG and wild-type MTG) to a constant activity level is performed by adding trypsin at 30°C Streptomyces hygroscopicus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.16
-
purified recombinant wild-type enzyme, uncleaved, pH 6.0, 37°C Streptomyces hygroscopicus
0.22
-
purified recombinant truncated enzyme mutant, uncleaved, pH 6.0, 37°C Streptomyces hygroscopicus
0.54
-
purified recombinant wild-type enzyme, cleaved by trypsin, pH 6.0, 37°C Streptomyces hygroscopicus
0.69
-
purified recombinant truncated enzyme mutant, cleaved by trypsin, pH 6.0, 37°C Streptomyces hygroscopicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
N-CBZ-Gln-Gly + hydroxylamine
-
Streptomyces hygroscopicus L-CBZ-Gln(gamma-monohydroxamate)-Gly + NH3
-
?
N-CBZ-Gln-Gly + hydroxylamine
-
Streptomyces hygroscopicus H197 L-CBZ-Gln(gamma-monohydroxamate)-Gly + NH3
-
?
protein glutamine + alkylamine
-
Streptomyces hygroscopicus protein N5-alkylglutamine + NH3
-
?
protein glutamine + alkylamine
-
Streptomyces hygroscopicus H197 protein N5-alkylglutamine + NH3
-
?

Synonyms

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

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
Streptomyces hygroscopicus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
thermal inactivation of MTG: the residual activity is measured after the purified MTG is incubated at 60°C for 30 min in the presence of glycerol, sorbitol, mannitol, sucrose, cellulose, and PEG, respectively. About 60% activity of the mutant MTG and about 70% activity of the wild-type MTG are lost. At the given concentrations of different stabilizers, glycerol and sorbitol exhibit a higher stabilizing effect on the mutant MTG and more than 60% activity of the mutant MTG is preserved, while sorbitol and mannitol present a higher stabilizing effect on the wild-type MTG and about 60% activity of the wild MTG is retained Streptomyces hygroscopicus
40
-
wild-type and truncated mutant MTGs show higher stability at 40°C and 1-3% salinity Streptomyces hygroscopicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 8
-
Streptomyces hygroscopicus

General Information

General Information Comment Organism
additional information the MTG has a triplet active center Cys151-Asp342-His361 Streptomyces hygroscopicus