Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | the inactive proform of the enzyme is activated by cleavage through trypsin | Streptomyces hygroscopicus |
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 | 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 | 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 | 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 | 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 | 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 | 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces hygroscopicus | A0A291LTQ0 | - |
- |
Streptomyces hygroscopicus H197 | A0A291LTQ0 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
proteolytic modification | the inactive proform of the enzyme is activated by cleavage through trypsin | Streptomyces hygroscopicus |
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 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 | 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 | Comment | Organism |
---|---|---|
microbial transglutaminase | - |
Streptomyces hygroscopicus |
MTG | - |
Streptomyces hygroscopicus |
transglutaminase | - |
Streptomyces hygroscopicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
- |
Streptomyces hygroscopicus |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | 8 | - |
Streptomyces hygroscopicus |
General Information | Comment | Organism |
---|---|---|
additional information | the MTG has a triplet active center Cys151-Asp342-His361 | Streptomyces hygroscopicus |