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Information on EC 2.3.2.13 - protein-glutamine gamma-glutamyltransferase and Organism(s) Streptomyces mobaraensis and UniProt Accession P81453
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2.3.2.13 protein-glutamine gamma-glutamyltransferaseIUBMB Comments
Requires Ca2+. The gamma-carboxamide groups of peptide-bound glutamine residues act as acyl donors, and the 6-amino-groups of protein- and peptide-bound lysine residues act as acceptors, to give intra- and inter-molecular N6-(5-glutamyl)-lysine crosslinks. Formed by proteolytic cleavage from plasma Factor XIII
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Word Map
- 2.3.2.13
- celiac
- coagulation
- clot
- fibrinogen
-
crosslinking
- gluten
-
serological
- bleeding
- thrombin
- autoimmune
- children
- autoantibody
- platelet
-
gluten-free
- keratinocytes
-
anti-tissue
- gliadin
- collagen
- duodenal
- igg
- bowel
-
deamidated
- fibronectin
-
villous
- keratin
-
fibrinolysis
- plasminogen
-
isopeptide
- involucrin
- enteropathy
- prothrombin
-
cornified
- plasmin
-
hemostatic
-
endoscopy
-
marsh
-
gelation
-
texture
- ichthyosis
-
fibrinopeptide
-
cryoprecipitate
-
gastroenterology
-
rheological
- leiden
-
malabsorption
- filaggrin
- nutrition
- analysis
- agriculture
- diagnostics
-
antifibrinolytic
- medicine
- synthesis
- food industry
- biotechnology
- histiocytomas
-
sealant
-
2-antiplasmin
The taxonomic range for the selected organisms is: Streptomyces mobaraensis
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
transglutaminase, factor xiii, tissue transglutaminase, tgase, factor xiiia, transglutaminase 2, microbial transglutaminase, mtgase, tgase 2, protein 4.2, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
microbial transglutaminase
-
MTG
-
factor XIIIa
-
-
-
-
fibrin stabilizing factor
-
-
-
-
fibrinoligase
-
-
-
-
glutaminylpeptide gamma-glutamyltransferase
-
-
-
-
glutamyltransferase, glutaminylpeptide gamma-
-
-
-
-
microbial transglutaminase
MTGase
polyamine transglutaminase
-
-
-
-
protein-glutamine gamma-glutamyltransferase
R-glutaminyl-peptide:amine gamma-glutamyl transferase
-
-
-
-
tissue transglutaminase
-
-
-
-
transglutaminase
transglutaminase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
protein glutamine + alkylamine = protein N5-alkylglutamine + NH3
ping-pong mechanism
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
aminoacyl group transfer
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
protein-glutamine:amine gamma-glutamyltransferase
Requires Ca2+. The gamma-carboxamide groups of peptide-bound glutamine residues act as acyl donors, and the 6-amino-groups of protein- and peptide-bound lysine residues act as acceptors, to give intra- and inter-molecular N6-(5-glutamyl)-lysine crosslinks. Formed by proteolytic cleavage from plasma Factor XIII
CAS REGISTRY NUMBER
COMMENTARY
80146-85-6
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
alpha-casein glutamine + alkylamine
alpha-casein N5-alkylglutamine + NH3
-
-
-
?
beta-casein glutamine + hydroxylamine
beta-casein N5-hydroxylglutamine + NH3
-
-
-
?
Cbz-Gln-Gly + D-lysine
Cbz-N5-aminocaproyl-glutaminyl-Gly + NH3
-
-
-
?
Cbz-Gln-Gly + hydroxylamine
Cbz-N5-hydroxyglutaminyl-Gly + NH3
-
-
-
?
Cbz-Gln-Gly + L-lysine
Cbz-N5-aminocaproyl-glutaminyl-Gly + NH3
-
-
-
?
CBZ-Gln-Gly-OH + hydroxylamine
CBZ-Gln(gamma-monohydroxamate)-Gly + NH3
-
-
-
?
fluorescein-4-isothiocyanate-beta-AQG + NK6-AP
fluorescein-4-isothiocyanate-labeled NK6-AP + ?
NK6-AP, recombinant Escherichia coli alkaline phosphatase with a N-terminal fused acyl-acceptor substrate peptide tag MKHKGS
-
-
?
fluorescein-4-isothiocyanate-epsilon-aminocaproate-QG + NK6-AP
fluorescein-4-isothiocyanate-labeled NK6-AP + ?
NK6-AP, recombinant Escherichia coli alkaline phosphatase with a N-terminal fused acyl-acceptor substrate peptide tag MKHKGS
-
-
?
N-CBZ-Glu-Gly + hydroxylamine
CBZ-Glu-(gamma-monohydroxamate)-Gly + NH3
-
-
-
?
sulforhodamine-beta-AQG + NK6-AP
sulforhodamine-labeled NK6-AP + ?
NK6-AP, recombinant Escherichia coli alkaline phosphatase with a N-terminal fused acyl-acceptor substrate peptide tag MKHKGS
-
-
?
Z-Gln-Gly + hydroxylamine
Z-N5-hydroxyglutaminyl-Gly + NH3
-
-
-
?
1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N-(5'-N',N'-dimethylamino-1'-naphthalenesulfonyl)diamidopentane + alpha-carbobenzoxy-lysine
?
-
fluorescent substrate for detection and characterization of glutamine acceptor compounds
-
-
?
1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N-(5'-N',N'-dimethylamino-1'-naphthalenesulfonyl)diamidopentane + alpha-S1-casein
?
-
fluorescent substrate for detection and characterization of glutamine acceptor compounds
-
-
?
1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N-(5'-N',N'-dimethylamino-1'-naphthalenesulfonyl)diamidopentane + butylamine
?
-
fluorescent substrate for detection and characterization of glutamine acceptor compounds
-
-
?
1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N-(5'-N',N'-dimethylamino-1'-naphthalenesulfonyl)diamidopentane + ethylamine
?
-
fluorescent substrate for detection and characterization of glutamine acceptor compounds
-
-
?
1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N-(5'-N',N'-dimethylamino-1'-naphthalenesulfonyl)diamidopentane + propylamine
?
-
fluorescent substrate for detection and characterization of glutamine acceptor compounds
-
-
?
Cbz-Gln-Gly + hydroxylamine
Cbz-Gln(gamma-monohydroxamate)-Gly + NH3
-
-
-
?
N-carbobenzoxy-L-glutaminylglycine + NH2OH
hydroxamic acid + ?
-
-
-
-
?
Nalpha-benzyloxycarbonyl-L-glutaminylglycine + hydroxylamine
?
-
-
-
-
?
putrescine-alginate conjugate + dimethylated casein
?
-
putrescine (1,4-diaminobutane) covalently linked to alginate and low-methoxyl pectin, although the latter at higher concentrations, are able to act as effective acyl acceptor transglutaminase substrates in vitro using both dimethylated casein and soy flour proteins as acyl donors
-
-
?
putrescine-pectin conjugate + dimethylated casein
?
-
putrescine (1,4-diaminobutane) covalently linked to alginate and low-methoxyl pectin, although the latter at higher concentrations, are able to act as effective acyl acceptor transglutaminase substrates in vitro using both dimethylated casein and soy flour proteins as acyl donors
-
-
?
putrescine-pectin conjugate + soy flour protein
?
-
-
reacion produces edible films with low water vapor permeability and improved mechanical properties
-
?
Streptomyces subtilisin and TAMEP inhibitor (SSTI) + N-lauroylsarcosine
?
-
TGase mediated biotinylation
-
-
?
YELQRPYHSELP + biotinylated cadaverine
?
-
preferred substrate, acitve even in the peptide form
-
-
?
YELQRPYHSELP-glutathione-S-transferase + biotinylated cadaverine
?
-
preferred substrate
-
-
?
Z-Gln-Gly + hydroxamate
L-glutamic acid gamma-monohydroxamate + NH3
-
high activity
-
-
?
Z-Gln-Gly + Nalpha-acetyl-L-lysine methyl ester
?
-
high activity
-
-
?
Z-Gln-Gly + propargylamine
Z-Nepsilon-propargyl-Gln-Gly + NH3
-
high activity
-
-
?
protein glutamine + alkylamine
protein N5-alkylglutamine + NH3
-
-
-
?
protein glutamine + alkylamine
protein N5-alkylglutamine + NH3
-
-
-
-
?
protein glutamine + alkylamine
protein N5-alkylglutamine + NH3
-
-
-
?
protein-bound gamma-glutamine + alkylamine
protein N5-alkylglutamine + NH3
-
-
-
-
?
protein-bound gamma-glutamine + alkylamine
protein N5-alkylglutamine + NH3
-
aliphatic amine donors incorporated into benzyloxycarbonyl-L-Gln-Gly: hydroxylamine, methylamine, ethylamine, n-propylamine, n-butylamine, n-pentylamine, n-hexylamine, amino acids incorporated: L-lysine and D-lysine, amino acid esters incorporated: Gly, Ala, Val, and Met ethyl esters, Lys-analogs incorporated: L-ornithine, aliphatic amines with omega-carboxyl groups incorporated: 5-aminovaleric acid, epsilon-amino-n-caproic acid, 7-aminoheptanoic acid, omega-aminocaprylic acid, amines with functional groups incorporated: carbonyl, phosphate, sulfo groups and saccharides
-
?
[protein]-L-glutamine + alkylamine
[protein]-N5-alkyl-L-glutamine + NH3
-
-
-
-
?
[protein]-L-glutamine + alkylamine
[protein]-N5-alkyl-L-glutamine + NH3
-
-
-
?
additional information
?
-
MTG can accept diverse fluorophores such asdansyl, fluorescein, and rhodamine derivatives in place of the benzyloxycarbonyl moiety when linked via a beta-alanine or epsilon-aminocaproic acid linker
-
-
?
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
-
-
-
additional information
?
-
crosslinking activity and IgG reactivity after digestion with cow and horse milk proteins, detailed overview
-
-
-
additional information
?
-
microbial transglutaminase (MTG) is a practical tool to enzymatically form isopeptide bonds between peptide or protein substrates. Engineered, highly reactive substrates of microbial transglutaminase enable protein labeling within various secondary structure elements. MTG can react readily with glutamines in alpha-helical, beta-sheet, and unstructured loop elements and does not favor one type of secondary structure. Building of a GB1 library where each variant contains a single glutamine at positions covering all secondary structure elements, detailed overview. The most reactive and selective variants display an over 100fold increase in incorporation compared to another developed aminated benzo[a]imidazo[2,1,5-cd]indolizine-type fluorophore, relative to native GB1
-
-
-
additional information
?
-
site-specific conjugation to native and engineered lysines in human immunoglobulins by microbial transglutaminase, overview. ESI-MS analysis of antibodies incubated with an acyl donor substrate and enzyme MTG, performed by incubation with ZQG-biotin and MTG at 37°C overnight followed by digestion with IdeS to generate Fab'2 and Fc fragments. A positive-control peptide with two known lysine acyl acceptor sites (GGSTKHKIPGGS) is genetically fused to the C-terminus of mAb1 HC or LC (HC-KTag or LC-KTag, respectively) and analyzed for transamidation. The addition of the KTag to the HC C-terminus blocks removal of Lys447, thereby allowing MTG to utilize Lys447 as an acyl acceptor site. Effect of single C-terminal amino acids on transamidation of HC Lys447, and analysis of transamidation of single lysine substitutions in gamma, iota, kappa, and lambda constant regions, overview. Optimal transamidation of an LC C-terminal lysine requires a spacer between Cys214 and the lysine. Analysis of transamidation of select single lysine substitutions, conducted by incubating samples with ZQG-biotin and MTG at 37°C overnight. Mutational analysis of binding sites
-
-
-
additional information
?
-
site-specific transglutaminase-mediated conjugation of interferon alpha-2b at glutamine or lysine residues. Reactivity of IFN alpha-2b to microbial transglutaminase (TGase) allows site-specific conjugation of this protein drug. Production of two monoderivatized isomers of IFN with high yields, mass spectrometry analysis of the two conjugates indicating that they are exclusively modified at the level of Gln101 if the protein is reacted in the presence of an amino-containing ligand (i.e. dansylcadaverine) or at the level of Lys164 if a glutamine-containing molecule is used (i.e. carbobenzoxy-L-glutaminyl-glycine, ZQG). The enzyme is absolutely specific, among the 10 Lys and 12 Gln residues of the protein, only Gln101 and Lys164 are located in highly flexible protein regions
-
-
-
additional information
?
-
SM-TAP procession of the pro-form zymogen is not essential for activity as TAMEP-treated and fully processed enzyme and the zymogen exhibit similar catalytic activity
-
-
-
additional information
?
-
-
SM-TAP procession of the pro-form zymogen is not essential for activity as TAMEP-treated and fully processed enzyme and the zymogen exhibit similar catalytic activity
-
-
-
additional information
?
-
TGase activity on the productivity of crosslinking peptide with tryptic casein, substrate specificity of wild-type and mutant enzymes, overview
-
-
-
additional information
?
-
-
TGase activity on the productivity of crosslinking peptide with tryptic casein, substrate specificity of wild-type and mutant enzymes, overview
-
-
-
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
-
-
-
additional information
?
-
the enzyme also catalyzes the deamination of amines. Glutamine is recognised as the acyl donor substrate by TGases due to the gamma-carboxyamide group. N-Benzyloxycarbonyl-L-glutaminylglycine (CBZ-Gln-Gly) is the standard glutamine peptide substrate used for TGases. Acyl acceptor substrates and acyl donor substrates, overview. The mTGase enzyme was reported to recognise L-isomer of lysine slightly more than its D-isomer when incorporated into a Z-Gln-Gly motif
-
-
-
additional information
?
-
-
posttranslational dimerization and multimerization of Camelidae anti-human TNF single domain antibodies in vitro catalyzed by microbial transglutaminases. Ribonuclease S-tag-peptide acts as a peptidyl substrate in covalent protein cross-linking reactions catalyzed by MTG. C-terminally fusion of the S-tag sequence to the anti-hTNF-variable heavy chain-domain results in fusion proteins that are efficiently dimerized and multimerized by MTG whereas anti-hTNF-variable heavy chain domain is not susceptible to protein crosslinking
-
-
?
additional information
?
-
-
no activity with carbamic acid, thiamine, 2-bromoethylamine, N-ethylmethylamine, sarcosine, butanol, butanethiol, or L-isoleucine methyl ester
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
alpha-casein glutamine + alkylamine
alpha-casein N5-alkylglutamine + NH3
-
-
-
?
Nalpha-benzyloxycarbonyl-L-glutaminylglycine + hydroxylamine
?
-
-
-
-
?
protein-bound gamma-glutamine + alkylamine
protein N5-alkylglutamine + NH3
-
-
-
-
?
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
-
-
-
protein glutamine + alkylamine
protein N5-alkylglutamine + NH3
-
-
-
?
protein glutamine + alkylamine
protein N5-alkylglutamine + NH3
-
-
-
-
?
protein glutamine + alkylamine
protein N5-alkylglutamine + NH3
-
-
-
?
[protein]-L-glutamine + alkylamine
[protein]-N5-alkyl-L-glutamine + NH3
-
-
-
-
?
[protein]-L-glutamine + alkylamine
[protein]-N5-alkyl-L-glutamine + NH3
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
NaCl
purification of a high-salt resistent microbial transglutaminase, isozyme MTG-TX, 79.8% activity remaining
additional information
additional information
the enzyme is not affected by Cu2+ and Fe3+
additional information
-
the enzyme is not affected by Ca2+, Ba2+, K+, or Na+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Monodansylcadaverine
-
inhibition of glutamyl transfer to putrescine-pectin and putrescine-alginate
additional information
isozyme MTG-TX is not inhibited by Li+, Na+, K+, Ca2+, Mg2+, Ba2+, Mn2+, or the inhibitor phenylmethanesulfonyl fluoride (PMSF)
-
additional information
-
heat-treated pro-enzyme acts effectively as inhibitor of mature form
-
additional information
-
not inhibitory: Ba2+, Ca2+, Co2+, Cu2+, Fe2+, Fe3+, Mg2+, Mn2+, Na+, phenylmethylsulfonyl fluoride, EDTA
-
additional information
-
enzyme precursor protein is inhibitory to mature enzyme, even after heat treatment. Precursor is secreted to culture medium
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
the enzyme is secreted as a pro-enzyme which is activated by the endoprotease TAMEP
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
52.66
Nalpha-benzyloxycarbonyl-L-glutaminylglycine
-
in 250 mM Tris acetate buffer (pH 8.0), at 37°C
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
-
8.55
CBZ-Gln-Gly-OH
recombinant wild-type enzyme, pH 6.0, 37°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00035
alpha-carbobenzoxy-lysine
-
1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N-(5'-N'-N'-dimethylamino-1'naphthalenesulfonyl)diamidopentane as glutamine donor
0.00055
Butylamine
-
1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N-(5'-N',N'-dimethylamino-1'-naphthalenesulfonyl)diamidopentane as glutamine donor
0.00055
ethylamine
-
1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N-(5'-N',N'-dimethylamino-1'-naphthalenesulfonyl)diamidopentane as glutamine donor
35.48
Nalpha-benzyloxycarbonyl-L-glutaminylglycine
-
in 250 mM Tris acetate buffer (pH 8.0), at 37°C
0.0004
Propylamine
-
1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N-(5'-N',N'-dimethylamino-1'-naphthalenesulfonyl)diamidopentane as glutamine donor
27.37
CBZ-Gln-Gly-OH
recombinant wild-type enzyme, pH 6.0, 37°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.67
Nalpha-benzyloxycarbonyl-L-glutaminylglycine
-
in 250 mM Tris acetate buffer (pH 8.0), at 37°C
3.2
CBZ-Gln-Gly-OH
recombinant wild-type enzyme, pH 6.0, 37°C
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
55.7
purified recombinant enzyme penta-site mutant DM01, pH and temperature not specified in the publication
30.5
30.6
32.3
32.4
35.5
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4 - 9
activity range, wild-type and mutant enzyme, profiles overview
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
20 - 60
activity range, wild-type and mutant enzyme, profiles overview
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
the immature enzyme is secreted, and extracellulary, the zymogen is processed by an endogenous metalloprotease called TGase-activating protease (TAMEP) and a tripeptidyl aminopeptidase (SM-TAP)
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
the removal of the pro-sequence region using proteases results in the active site of the mature enzyme to be revealed to initiate the reaction
physiological function
additional information
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
physiological function
microbial transglutaminase alters the immunogenic potential and cross-reactivity of horse and cow milk proteins. The effect of TG on immunoreactivity depends on enzyme quantity and milk protein type. Determination of immunoreactivity of milk proteins by competitive ELISA
additional information
comparison of the different TGases from Streptomyces mobaranensis and Streptomyces cinnamoneus, and of a chimeric mutant constructed from both, overview
additional information
-
comparison of the different TGases from Streptomyces mobaranensis and Streptomyces cinnamoneus, and of a chimeric mutant constructed from both, overview
additional information
effect of microbial transglutaminase on the mechanical properties and microstructure of acid-induced gels and emulsion gels produced from thermal denatured egg white proteins (TD-EWPs), overview. Impact of TGase on mechanical, rheological, and microstructural properties of cold-set EWP gels and emulsion gels produced from the TD-EWP. EWP is sufficiently cross-linked by MTGase
additional information
mechanism of mTG-catalyzed isopeptide bond formation between protein-bound glutamine and lysine residues. Structure-function analysis, substrate specificity, overview
additional information
-
mechanism of mTG-catalyzed isopeptide bond formation between protein-bound glutamine and lysine residues. Structure-function analysis, substrate specificity, overview
additional information
mTGase structure-function relationship, overview. The active site is a single cysteine (C64) residing at the bottom cleft of the crystal structure (PDB ID 3IU0) where it forms a catalytic triad with the aspartic acid (D255) and histidine (H274) residues. The pro-sequence region is vital for enzyme folding and inhibition of enzyme activation within the cells to avoid detrimental cross-linking of cytosolic proteins. The enzyme structure has a wide active site cleft position that accommodates the alpha-helix pro-sequence. This unique attribute explains the broad substrate specificity for acyl donors which allows for additional flexibility in the active site to accommodate a less specific substrate, overview
additional information
the enzyme shows a ping-pong mechanism
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
42500
45000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
proteolytic modification
proteolytic modification
Streptomyces TGase is secreted as a zymogen with an additional prosequence at the N-terminus, the cleavage of the prosequence activates the enzyme
proteolytic modification
the immature enzyme is secreted, and extracellulary, the zymogen is processed by an endogenous metalloprotease called TGase-activating protease (TAMEP) and a tripeptidyl aminopeptidase (SM-TAP). SM-TAP procession is not essential for activity as TAMEP-treated and fully processed zymogen exhibits similar catalytic activity
proteolytic modification
-
pro-enzyme acts effectively as inhibitor of mature form
proteolytic modification
-
enzyme precursor protein is inhibitory to mature enzyme, even after heat treatment. Precursor is secreted to culture medium
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
docking model of MTG binding to substrate N-carbobenzoxy-L-glutaminyl-glycine. Substrate is stretched along the MTG active site cleft with hydrophobic and/or aromatic residues interacting directly with the substrate. An oxyanion binding site for TGase activity may be constructed from the amide groups of Cys64 and/or Val65
hanging drop vapor diffusion method, using 30% (w/v) PEG 8000, 50 mM NaCl, 1 mM EDTA, 1 mM 2-mercaptoethanol, 0.01% (w/v) NaN3, 100 mM cacodylic acid (pH 5.0), and 2% (v/v) glycerol
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C64A
D255A
E164L
site-directed mutagenesis, the E164L mutant exhibits a 1.95fold increased specific activity and 1.66fold increased half-life at 50°C compared to wild-type. The molecular dynamics (MD) simulation results indicate that the mutation Glu164Leu results in weaker interactions of Asp159-Glu164 and Gly228-Leu231, leading to the enhanced instability of Ile240-Asn253 linked to Gly228-Leu231 by eight residues. It further causes reduced interactions between loop region 1 (Ile240-Asn253) and loop region 2 (His277-Met288), facilitating the access of substrate molecule to the active site. Structure-activity relationship for MTG adapted to high temperature conditions. Enhancing activity and thermostability of Streptomyces mobaraensis transglutaminase by directed evolution, Molecular mechanism of improved activity of E164L analyzed by molecular dynamics simulations
N253A
S2P
site-directed mutagenesis, the mutant shows increased activity compared to wild-type
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
Y256A
A10S
-
the mutant shows higher specific activity compared to the wild type enzyme
D14N
-
the mutant shows higher specific activity compared to the wild type enzyme
D20A
-
the mutant shows reduced activity compared to the wild type enzyme
D301A
-
the mutation drastically reduces the catalytic activity of the enzyme
D3F
-
the mutant shows higher specific activity compared to the wild type enzyme
D3L
-
the mutant shows higher specific activity compared to the wild type enzyme
D3N
-
the mutant shows higher specific activity compared to the wild type enzyme
E28D
-
the mutant shows higher specific activity compared to the wild type enzyme
E29A
-
the mutant shows about 60% reduced activity compared to the wild type enzyme
E58D
-
the mutant shows higher specific activity compared to the wild type enzyme
H289F
-
the mutant shows higher specific activity compared to the wild type enzyme
H289Y
-
the mutant shows higher specific activity compared to the wild type enzyme
I24A
-
the mutant shows reduced activity compared to the wild type enzyme
L27A
-
the mutant shows about 40% reduced activity compared to the wild type enzyme
M16T
-
the mutant shows higher specific activity compared to the wild type enzyme
M16T/G283S
-
the mutant shows higher specific activity compared to the wild type enzyme
N23A
-
the mutant shows about 50% reduced activity compared to the wild type enzyme
N25A
-
the mutant shows reduced activity compared to the wild type enzyme
N28A
-
the mutant shows about 45% reduced activity compared to the wild type enzyme
N320D
-
the mutant shows higher specific activity compared to the wild type enzyme
N32D
-
the mutant shows higher specific activity compared to the wild type enzyme
N32D/E264D/N320T
-
the mutant shows higher specific activity compared to the wild type enzyme
P12S
-
the mutant shows higher specific activity compared to the wild type enzyme
Q74A
-
the mutant shows higher specific activity compared to the wild type enzyme
Q74L
-
the mutant shows higher specific activity compared to the wild type enzyme
Q74N
-
the mutant shows higher specific activity compared to the wild type enzyme
R238F
-
the mutant shows higher specific activity compared to the wild type enzyme
R238L
-
the mutant shows higher specific activity compared to the wild type enzyme
R26F
-
the mutant shows higher specific activity compared to the wild type enzyme
R26L
-
the mutant shows higher specific activity compared to the wild type enzyme
R5K
-
the mutant shows higher specific activity compared to the wild type enzyme
S199A
-
the mutant shows higher specific activity compared to the wild type enzyme
S284T
-
the mutant shows higher specific activity compared to the wild type enzyme
S299L
-
the mutant shows higher specific activity compared to the wild type enzyme
S303A
-
the mutant shows higher specific activity compared to the wild type enzyme
S303F
-
the mutant shows higher specific activity compared to the wild type enzyme
S303T
-
the mutant shows higher specific activity compared to the wild type enzyme
T77A
-
the mutant shows higher specific activity compared to the wild type enzyme
T77F
-
the mutant shows higher specific activity compared to the wild type enzyme
T77L
-
the mutant shows higher specific activity compared to the wild type enzyme
T77S
-
the mutant shows higher specific activity compared to the wild type enzyme
V30D
-
the mutant shows higher specific activity compared to the wild type enzyme
V30I
-
the mutant shows higher specific activity compared to the wild type enzyme
V30T
-
the mutant shows higher specific activity compared to the wild type enzyme
V65I
-
the mutant shows higher specific activity compared to the wild type enzyme
V6T
-
the mutant shows higher specific activity compared to the wild type enzyme
W59F
-
the mutant shows higher specific activity compared to the wild type enzyme
Y10A
-
the mutant shows reduced activity compared to the wild type enzyme
Y34F
-
the mutant shows higher specific activity compared to the wild type enzyme
Y34F/D268N
-
the mutant shows higher specific activity compared to the wild type enzyme
Y42H
-
the mutant shows higher specific activity compared to the wild type enzyme
Y75A
-
the mutant shows higher specific activity compared to the wild type enzyme
Y75F
-
the mutant shows higher specific activity compared to the wild type enzyme
Y75H
-
the mutant shows higher specific activity compared to the wild type enzyme
additional information
additional information
construction of a chimeric mutant constructed from the TGases of Streptomyces mobaranensis (SMTG) and Streptomyces cinnamoneus (SCTG), the mutant enzyme consists of the N-terminal half of SCTG and the C-terminal half of SMTG
additional information
-
construction of a chimeric mutant constructed from the TGases of Streptomyces mobaranensis (SMTG) and Streptomyces cinnamoneus (SCTG), the mutant enzyme consists of the N-terminal half of SCTG and the C-terminal half of SMTG
additional information
development and evaluation of a method for production of a reusable immobilized recombinant His-tagged Escherichia coli biotin ligase (BirA) onto amine-modified magnetic microspheres (MMS) via covalent cross-linking catalyzed using microbial transglutaminase (MTG). The site-specifically immobilized BirA exhibited approximately 95% of enzymatic activity of the free BirA, and without a significant loss in intrinsic activity after 10 rounds of recycling. Method, overview
additional information
development of enzyme engineering to improve, alter, or customise the functional properties of mTGase, e.g. thermoengineering for better heat stability and heat sensitivity, overview. The N-termius of mTGase is an important region that influences the thermal properties of the enzyme due to the fact that all single-point mutations related to the altered thermal properties are located in this area, random mutagenesis. Semirational mutagenesis is also successful to isolate mTGase variants with increased thermostabilities. Seven hot spot residues, which are reported to be the thermostabilizing sites, are mutated for the generation of mutant libraries to screen for thermostable variants. Later, variants with single amino acid substitution comprising of the highest thermostabilities are mixed by DNA shuffling to generate a secondary library for screening. Finally, the variants with improved thermostabilities are isolated via standard assay. For production of soluble enzyme, introduction of a fusion partner with the extension of the N-terminal region to contain few LacZ residues followed by the first 20 residues of enzyme purine nucleoside phosphorylase is done. This strategy results in the accumulation of high levels of mTGase in the cytoplasm. The thermoinducible expression system yields a lower protein yield but produces the enzyme with a higher specificity as no major modification is done to the enzyme making it preferable compared to constitutive expression system
additional information
effect of microbial transglutaminase on the mechanical properties and microstructure of acid-induced gels and emulsion gels produced from thermal denatured egg white proteins. Impact of TGase on mechanical, rheological, and microstructural properties of cold-set EWP gels and emulsion gels produced from the TD-EWP, preparation of cold-set TD-EWP emulsion gels reinforced with MTGase, size determination of emulsion droplets and evaluation of MTGase mediated-covalent cross-linking in gels and emulsion gels by SDS-PAGE, method, overview
additional information
engineering an automaturing transglutaminase with enhanced thermostability by genetic code expansion with two codon reassignments. The first amino acid, 3-chloro-L-tyrosine, is incorporated into microbial transglutaminase (MTG) in response to in-frame UAG codons to substitute for the 15 tyrosine residues separately. The two substitutions at positions 20 and 62 are found to each increase thermostability of the enzyme, while the seven substitutions at positions 24, 34, 75, 146, 171, 217, and 310 exhibit neutral effects. Then, these two stabilizing chlorinations are combined with one of the neutral ones, and the most stabilized variant is found to contain 3-chlorotyrosines at positions 20, 62, and 171, exhibiting a half-life 5.1fold longer than that of the wild-type enzyme at 60°C. Next, this MTG variant is further modified by incorporating the alpha-hydroxy acid analogue of Nepsilon-allyloxycarbonyl-L-lysine (AlocKOH), specified by the AGG codon, at the end of the N-terminal inhibitory peptide. The ester bond, thus incorporated into the main chain, efficiently self-cleaves under alkaline conditions (pH 11.0), achieving the autonomous maturation of the thermostabilized MTG in transformed Escherichia coli. Method, overview
additional information
-
engineering an automaturing transglutaminase with enhanced thermostability by genetic code expansion with two codon reassignments. The first amino acid, 3-chloro-L-tyrosine, is incorporated into microbial transglutaminase (MTG) in response to in-frame UAG codons to substitute for the 15 tyrosine residues separately. The two substitutions at positions 20 and 62 are found to each increase thermostability of the enzyme, while the seven substitutions at positions 24, 34, 75, 146, 171, 217, and 310 exhibit neutral effects. Then, these two stabilizing chlorinations are combined with one of the neutral ones, and the most stabilized variant is found to contain 3-chlorotyrosines at positions 20, 62, and 171, exhibiting a half-life 5.1fold longer than that of the wild-type enzyme at 60°C. Next, this MTG variant is further modified by incorporating the alpha-hydroxy acid analogue of Nepsilon-allyloxycarbonyl-L-lysine (AlocKOH), specified by the AGG codon, at the end of the N-terminal inhibitory peptide. The ester bond, thus incorporated into the main chain, efficiently self-cleaves under alkaline conditions (pH 11.0), achieving the autonomous maturation of the thermostabilized MTG in transformed Escherichia coli. Method, overview
additional information
introducing point mutations within MTG's active site increases reactivity toward the most reactive substrate variant, I6Q-GB1, enhancing MTG's capacity to fluorescently label an engineered, highly reactive glutamine substrate
additional information
mTG crosslinked gelatin hydrogel preparation
additional information
mutiple-site mutagenesis of Streptomyces mobaraensis transglutaminase is performed in Escherichia coli. According to enzymatic assay and thermostability study, among three penta-site MTG mutants (DM01-03), DM01 exhibits the highest enzymatic activity of 55.7 U/mg and longest half-life at 50°C (418.2 min) and 60°C (24.8 min)
additional information
-
mutiple-site mutagenesis of Streptomyces mobaraensis transglutaminase is performed in Escherichia coli. According to enzymatic assay and thermostability study, among three penta-site MTG mutants (DM01-03), DM01 exhibits the highest enzymatic activity of 55.7 U/mg and longest half-life at 50°C (418.2 min) and 60°C (24.8 min)
additional information
peptidyl-linker sequences used that facilitate modification by microbial transglutaminase, overview
additional information
-
peptidyl-linker sequences used that facilitate modification by microbial transglutaminase, overview
additional information
site-specific transglutaminase-mediated conjugation of interferon alpha-2b at glutamine or lysine residues
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
additional information
utility of single lysine substitutions and the C-terminal Lys447 for engineering efficient acyl acceptor sites suitable for site-specific conjugation to a range of glutamine-based acyl donor substrates. Because recombinant mAbs lack the C-terminal Lys447 due to cleavage by carboxypeptidase B in the production cell host, it is analyzed if blocking the cleavage of Lys447 by the addition of a C-terminal amino acid can result in transamidation of Lys447 by a variety of acyl donor substrates. MTG efficiently transamidates Lys447 in the presence of any nonacidic, nonproline amino acid residue at position 448. Scanning mutagenesis of the hinge region in a Fab' fragment reveals sites of transamidation that are not reactive in the context of the full-length mAb. A positive-control peptide with two known lysine acyl acceptor sites (GGSTKHKIPGGS) is genetically fused to the C-terminus of mAb1 HC or LC (HC-KTag or LC-KTag, respectively) and analyzed for transamidation. The addition of the KTag to the HC C-terminus blocks removal of Lys447, thereby allowing MTG to utilize Lys447 as an acyl acceptor site. Mutational analysis of binding sites
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 10
the high-salt resistent isozyme MTG-TX exhibits a broad range of pH and temperature stability
759632
7
after treatment at 50°C and pH 7.0 for 60, 80, 100, and 120 min, respectively, recombinant mutant E164L maintains 60%, 41%, 29%, and 16% of its original activity
758740
5 - 9
-
the enzyme is stable within a pH range from 5.0 to 10.0 at 4°C for 12 h and within a pH range from 5.0 to 9.0 at 37°C for 30 min
720112
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50
60
additional information
the high-salt resistent isozyme MTG-TX exhibits a broad range of pH and temperature stability
50
after treatment at 50°C and pH 7.0 for 60, 80, 100, and 120 min, respectively, recombinant mutant E164L maintains 60%, 41%, 29%, and 16% of its original activity
50
purified recombinant enzyme penta-site mutant DM01, half-life is 418.2 min
60
purified recombinant enzyme penta-site mutant DM01, half-life is 24.8 min
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
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purification of extracellular high-salt resistent microbial transglutaminase, isozyme MTG-TX, by ethanol precipitation, cation exchange chromatography, hydrophobic interaction chromatography, and gel filtration, method optimization, overview
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21Gold (DE3) by nickel affinity chromatography
recombinant soluble enzyme from cell-free supernatant of Streptomyces lividans strain 1326 by ammonium sulfate fractionation, dialysis, and ultrafiltration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
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
recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
recombinant expression of engineered mutant enzyme in Escherichia coli
recombinant expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21Gold (DE3)
recombinant expression of wild-type and chimeric mutant enzymes in Streptomyces lividans strain 1326, Streptomyces TGase is secreted as a zymogen with an additional prosequence at the N-terminus
recombinant expression of wild-type and mutant enzymes in Corynebacterium glutamicum strain ATCC 13032, subcloning in Escherichia coli strain DH5alpha
recombinant expression of wild-type and mutant enzymes in Escherichia coli
expression in Escherichia coli as fusion protein with C-terminal His-tag
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
food industry
medicine
synthesis
biotechnology
food industry
-
enzyme-catalyzed cross-linking is effective in improving functional properties of stirred yak yoghurt (treated yoghurt produces a strong acid gel, higher consistency, cohesiveness, index of viscosity, and creamier mouth feel than the untreated product). Furtermore, enzyme-treated yak yoghurt presents lower wet yak hair or sweat odor, or both.
nutrition
synthesis
-
method for on-column activation of His-tagged enzyme by trypsin. About 89% of pro-MTG-His6 can be transferred to mature MTG-His6 with storage stabilization
additional information
microbial transglutaminase alters the immunogenic potential and cross-reactivity of horse and cow milk proteins. Possibility of reducing the immunoreactivity of horse milk proteins by microbial transglutaminase (TG) polymerization. The diet based on modified horse milk proteins could be an alternative for some patients with cow milk protein allergy
biotechnology
developments in mTGase engineering together with its role in biomedical applications including biomaterial fabrication for tissue engineering and biotherapeutics, overview
biotechnology
in production of homogeneous antibody-drug conjugates, the enzyme is useful for site-specific conjugation of glutamine-based acyl donor substrates and drugs to native and engineered lysines in human immunoglobulins by microbial transglutaminase, overview
biotechnology
microbial transglutaminase (mTG) is used as a crosslinking agent in the preparation of gelatin sponges. The physical properties of the materials are evaluated by measuring their material porosity, water absorption, and elastic modulus. The stability of the sponges are assessed via hydrolysis and enzymolysis, overview. To evaluate the cell compatibility of them TG crosslinked gelatin sponges (mTG sponges), adipose-derived stromal stem cells are cultured and inoculated into the scaffold. Cell proliferation and viability are measured using alamarBlue assay and LIVE/DEAD fluorescence staining, respectively. Cell adhesion on the sponges is observed by scanning electron microscopy. mTG sponges have uniform pore size, high porosity and water absorption, and good mechanical properties. In subcutaneous implantation (in Sprague-Dawley rats), the material is partially degraded in the first month and completely absorbed in the third month. Cell experiments show evident cell proliferation and high viability. The cells grow vigorously and adhered tightly to the sponge. In conclusion, mTG sponge has good biocompatibility and can be used in tissue engineering and regenerative medicine
biotechnology
the microbial transglutaminase is used for biotechnological and biomedical engineering, protein engineering by post-translational modification towards the generation of multifunctional conjugates. Biotechnological applications, detailed overview. Transglutaminase-mediated surface immobilization, a widely-used technique to increase stability of labile and cost-intensive enzymes and enable their reuse
biotechnology
use of MTG to site-specifically and covalently immobilize a substrate peptide-tagged protein, e.g. BirA, to a support, i.e. amine-modified magnetic microspheres (MMS)
food industry
microbial transglutaminase (MTG) is an enzyme widely used in the food industry
food industry
the enzyme is widely exploited in the meat processing industries. Enzyme mTGase is also widely applied in other food and textile industries by catalysing the formation of isopeptide bonds between peptides or protein substrates
food industry
transglutaminase MTG-TX can be used in food-related applications in salty environment, e.g. bacon or seafood
medicine
developments in mTGase engineering together with its role in biomedical applications including biomaterial fabrication for tissue engineering and biotherapeutics, biomedical engineering, overview
medicine
microbial transglutaminase (mTG) is used as a crosslinking agent in the preparation of gelatin sponges. The physical properties of the materials are evaluated by measuring their material porosity, water absorption, and elastic modulus. The stability of the sponges are assessed via hydrolysis and enzymolysis, overview. To evaluate the cell compatibility of them TG crosslinked gelatin sponges (mTG sponges), adipose-derived stromal stem cells are cultured and inoculated into the scaffold. Cell proliferation and viability are measured using alamarBlue assay and LIVE/DEAD fluorescence staining, respectively. Cell adhesion on the sponges is observed by scanning electron microscopy. mTG sponges have uniform pore size, high porosity and water absorption, and good mechanical properties. In subcutaneous implantation (in Spraguex15Dawley rats), the material is partially degraded in the first month and completely absorbed in the third month. Cell experiments show evident cell proliferation and high viability. The cells grow vigorously and adhered tightly to the sponge. In conclusion, mTG sponge has good biocompatibility and can be used in tissue engineering and regenerative medicine
medicine
the microbial transglutaminase is used for biotechnological and biomedical engineering, protein engineering by post-translational modification towards the generation of multifunctional conjugates. Biotechnological applications, detailed overview. Construction of protein-polymer and of antibody-drug conjugations for pharmaceutical applications
synthesis
microbial transglutaminase (MTG) is a practical tool to enzymatically form isopeptide bonds between peptide or protein substrates, crosslinking the side-chains of reactive glutamine and lysine residues is solidly rooted in food and textile processing. MTG-reactive glutamines can be readily introduced into a protein domain for fluorescent labeling, method evaluation, overview
synthesis
TGase can be used for the development of site-specific derivatives of IFN alpha-2b possessing interesting antiviral and pharmacokinetic properties
synthesis
the enzyme is used for biomaterial fabrication for tissue engineering, e.g. from gelatin, evaluation, overview
biotechnology
-
after fermentation in presence of enzyme, wheat dough has higher resistance to stretching and lower extensibility than control, dough contains more of the smallest and less large air bubbles. Enzyme improves formation of protein network in bread baked from normal or organic flour but at higher dosage causes uneven ditribution
biotechnology
-
reaction product of putrescine-pectin conjugate and soy flour protein may be used foredible films with low water vapor permeability and improved mechanical properties
nutrition
-
MTGase treatment significantly increases the denaturation temperature of beta-lactoglobulin in whey protein isolate, from 71.84°C in the untreated sample to 78.50°C after 30 h of incubation with MTGase. Increase in ´denaturation temperature is primarily due to covalent cross-linking and not due to an increase in nonpolar interactions within the protein. The surface hydrophobicity of the protein decreases upon cross-linking, due to occlusion of the hydrophobic cavities to the fluorescent probes. The cross-linked protein exhibits a U-shaped pH-stability profile with maximumturbidity at pH 4.0-4.5
nutrition
-
the functionality of light roasted peanut flour dispersions containing supplemental casein is altered after polymerization with microbial transglutaminase. The formation of high molecular weight covalent cross-links is observed. The gelling temperature of TGase-treated peanut flour dispersions containing 2.5% casein is significantly raised compared to the nontreated peanut flour-casein control solutions. The gel strength and water holding capacity of cross-linked peanut flour-casein test samples containing 5% casein is increased, while the yield stress and apparent viscosity are lowered compared to control dispersions. Casein is an effective cosubstrate with peanut flour for creating TGase-modified peanut flour-casein dispersions for use as a novel high protein food ingredient
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Pasternack, R.; Laurent, H.P.; Ruth, T.; Kaiser, A.; Schon, N.; Fuchsbauer, H.L.
A fluorescent substrate of transglutaminase for detection and characterization of glutamine acceptor compounds
Anal. Biochem.
249
54-60
1997
Cavia porcellus, Streptomyces mobaraensis
Ohtsuka, T.; Sawa, A.; Kawabata, R.; Nio, N.; Motoki, M.
Substrate specificities of microbial transglutaminase for primary amines
J. Agric. Food Chem.
48
6230-6233
2000
Cavia porcellus, Streptomyces mobaraensis
Autio, K.; Kruus, K.; Knaapila, A.; Gerber, N.; Flander, L.; Buchert, J.
Kinetics of transglutaminase-induced cross-linking of wheat proteins in dough
J. Agric. Food Chem.
53
1039-1045
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Streptomyces mobaraensis
Lu, S.Y.; Zhou, N.D.; Tian, Y.P.; Li, H.Z.; Chen, J.
Purification and properties of transglutaminase from Streptoverticillium mobaraense
J. Food Biochem.
27
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2003
Streptomyces mobaraensis
Pfleiderer, C.; Mainusch, M.; Weber, J.; Hils, M.; Fuchsbauer, H.L.
Inhibition of bacterial transglutaminase by its heat-treated pro-enzyme
Microbiol. Res.
160
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2005
Streptomyces mobaraensis
Schmidt, S.; Adolf, F.; Fuchsbauer, H.L.
The transglutaminase activating metalloprotease inhibitor from Streptomyces mobaraensis is a glutamine and lysine donor substrate of the intrinsic transglutaminase
FEBS Lett.
582
3132-3138
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Streptomyces mobaraensis, Streptomyces mobaraensis 40847
Di Pierro, P.; Mariniello, L.; Sorrentino, A.; Villalonga, R.; Chico, B.; Porta, R.
Putrescine-polysaccharide conjugates as transglutaminase substrates and their possible use in producing crosslinked films
Amino Acids
38
669-675
2010
Streptomyces mobaraensis
Sugimura, Y.; Yokoyama, K.; Nio, N.; Maki, M.; Hitomi, K.
Identification of preferred substrate sequences of microbial transglutaminase from Streptomyces mobaraensis using a phage-displayed peptide library
Arch. Biochem. Biophys.
477
379-383
2008
Streptomyces mobaraensis
Yang, H.L.; Pan, L.; Lin, Y.
Purification and on-column activation of a recombinant histidine-tagged pro-transglutaminase after soluble expression in Escherichia coli
Biosci. Biotechnol. Biochem.
73
2531-2534
2009
Streptomyces mobaraensis
Clare, D.A.; Gharst, G.; Maleki, S.J.; Sanders, T.H.
Effects of transglutaminase catalysis on the functional and immunoglobulin binding properties of peanut flour dispersions containing casein
J. Agric. Food Chem.
56
10913-10921
2008
Streptomyces mobaraensis
Agyare, K.K.; Damodaran, S.
pH-stability and thermal properties of microbial transglutaminase-treated whey protein isolate
J. Agric. Food Chem.
58
1946-1953
2010
Streptomyces mobaraensis
Plagmann, I.; Chalaris, A.; Kruglov, A.A.; Nedospasov, S.; Rosenstiel, P.; Rose-John, S.; Scheller, J.
Transglutaminase-catalyzed covalent multimerization of Camelidae anti-human TNF single domain antibodies improves neutralizing activity
J. Biotechnol.
142
170-178
2009
Streptomyces mobaraensis
Kamiya, N.; Abe, H.; Goto, M.; Tsuji, Y.; Jikuya, H.
Fluorescent substrates for covalent protein labeling catalyzed by microbial transglutaminase
Org. Biomol. Chem.
7
3407-3412
2009
Streptomyces mobaraensis (P81453)
Tagami, U.; Shimba, N.; Nakamura, M.; Yokoyama, K.; Suzuki, E.; Hirokawa, T.
Substrate specificity of microbial transglutaminase as revealed by three-dimensional docking simulation and mutagenesis
Protein Eng. Des. Sel.
22
747-752
2009
Streptomyces mobaraensis (P81453)
Yokoyama, K.; Utsumi, H.; Nakamura, T.; Ogaya, D.; Shimba, N.; Suzuki, E.; Taguchi, S.
Screening for improved activity of a transglutaminase from Streptomyces mobaraensis created by a novel rational mutagenesis and random mutagenesis
Appl. Microbiol. Biotechnol.
87
2087-2096
2010
Streptomyces mobaraensis, Streptomyces mobaraensis s-8112
Spolaore, B.; Raboni, S.; Ramos Molina, A.; Satwekar, A.; Damiano, N.; Fontana, A.
Local unfolding is required for the site-specific protein modification by transglutaminase
Biochemistry
51
8679-8689
2012
Streptomyces mobaraensis
Yang, M.T.; Chang, C.H.; Wang, J.M.; Wu, T.K.; Wang, Y.K.; Chang, C.Y.; Li, T.T.
Crystal structure and inhibition studies of transglutaminase from Streptomyces mobaraense
J. Biol. Chem.
286
7301-7307
2011
Streptomyces mobaraensis
Zhang, L.; Zhang, L.; Yi, H.; Du, M.; Ma, C.; Han, X.; Feng, Z.; Jiao, Y.; Zhang, Y.
Enzymatic characterization of transglutaminase from Streptomyces mobaraensis DSM 40587 in high salt and effect of enzymatic cross-linking of yak milk proteins on functional properties of stirred yogurt
J. Dairy Sci.
95
3559-3568
2012
Streptomyces mobaraensis, Streptomyces mobaraensis DSM 40587
Gundersen, M.T.; Keillor, J.W.; Pelletier, J.N.
Microbial transglutaminase displays broad acyl-acceptor substrate specificity
Appl. Microbiol. Biotechnol.
98
219-230
2014
Streptomyces mobaraensis
Salis, B.; Spinetti, G.; Scaramuzza, S.; Bossi, M.; Saccani Jotti, G.; Tonon, G.; Crobu, D.; Schrepfer, R.
High-level expression of a recombinant active microbial transglutaminase in Escherichia coli
BMC Biotechnol.
15
84
2015
Streptomyces mobaraensis (Q2VI01), Streptomyces mobaraensis
Rickert, M.; Strop, P.; Lui, V.; Melton-Witt, J.; Farias, S.E.; Foletti, D.; Shelton, D.; Pons, J.; Rajpal, A.
Production of soluble and active microbial transglutaminase in Escherichia coli for site-specific antibody drug conjugation
Protein Sci.
25
442-455
2016
Streptomyces mobaraensis
Ohtake, K.; Mukai, T.; Iraha, F.; Takahashi, M.; Haruna, K.I.; Date, M.; Yokoyama, K.; Sakamoto, K.
Engineering an automaturing transglutaminase with enhanced thermostability by genetic code expansion with two codon reassignments
ACS Synth. Biol.
7
2170-2176
2018
Streptomyces mobaraensis (P81453), Streptomyces mobaraensis
Boehme, B.; Moritz, B.; Wendler, J.; Hertel, T.C.; Ihling, C.; Brandt, W.; Pietzsch, M.
Enzymatic activity and thermoresistance of improved microbial transglutaminase variants
Amino Acids
52
313-326
2020
Streptomyces mobaraensis (P81453), Streptomyces mobaraensis
Yu, C.M.; Zhou, H.; Zhang, W.F.; Yang, H.M.; Tang, J.B.
Site-specific, covalent immobilization of BirA by microbial transglutaminase A reusable biocatalyst for invitro biotinylation
Anal. Biochem.
511
10-12
2016
Streptomyces mobaraensis (P81453)
Chan, S.K.; Lim, T.S.
Bioengineering of microbial transglutaminase for biomedical applications
Appl. Microbiol. Biotechnol.
103
2973-2984
2019
Streptomyces mobaraensis (P81453)
Liu, Y.; Huang, L.; Shan, M.; Sang, J.; Li, Y.; Jia, L.; Wang, N.; Wang, S.; Shao, S.; Liu, F.; Lu, F.
Enhancing the activity and thermostability of Streptomyces mobaraensis transglutaminase by directed evolution and molecular dynamics simulation
Biochem. Eng. J.
151
107333
2019
Streptomyces mobaraensis (P81453)
-
Spolaore, B.; Raboni, S.; Satwekar, A.A.; Grigoletto, A.; Mero, A.; Montagner, I.M.; Rosato, A.; Pasut, G.; Fontana, A.
Site-specific transglutaminase-mediated conjugation of interferon alpha-2b at glutamine or lysine residues
Bioconjug. Chem.
27
2695-2706
2016
Streptomyces mobaraensis (P81453)
Spidel, J.L.; Vaessen, B.; Albone, E.F.; Cheng, X.; Verdi, A.; Kline, J.B.
Site-specific conjugation to native and engineered lysines in human immunoglobulins by microbial transglutaminase
Bioconjug. Chem.
28
2471-2484
2017
Streptomyces mobaraensis (P81453)
Deweid, L.; Avrutina, O.; Kolmar, H.
Microbial transglutaminase for biotechnological and biomedical engineering
Biol. Chem.
400
257-274
2019
Bacillus subtilis (P40746), Streptomyces mobaraensis (P81453), Streptomyces mobaraensis, Kutzneria albida (W5WHY8), Bacillus subtilis 168 (P40746), Kutzneria albida DSM 43870 (W5WHY8)
Mu, D.; Lu, J.; Shu, C.; Li, H.; Li, X.; Cai, J.; Luo, S.; Yang, P.; Jiang, S.; Zheng, Z.
Improvement of the activity and thermostability of microbial transglutaminase by multiple-site mutagenesis
Biosci. Biotechnol. Biochem.
82
106-109
2018
Streptomyces mobaraensis (P81453), Streptomyces mobaraensis
Tokai, S.; Uraji, M.; Hatanaka, T.
Molecular insights into the mechanism of substrate recognition of Streptomyces transglutaminases
Biosci. Biotechnol. Biochem.
84
575-582
2020
Streptomyces mobaraensis (P81453), Streptomyces mobaraensis, Streptomyces cinnamoneus (Q8GR90), Streptomyces cinnamoneus, Streptomyces cinnamoneus NBRC 13864 (Q8GR90), Streptomyces mobaraensis NBRC 13819 (P81453)
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
BMC Biotechnol.
17
23
2017
Streptomyces mobaraensis (P81453)
Alavi, F.; Emam-Djomeh, Z.; Salami, M.; Mohammadian, M.
Effect of microbial transglutaminase on the mechanical properties and microstructure of acid-induced gels and emulsion gels produced from thermal denatured egg white proteins
Int. J. Biol. Macromol.
153
523-532
2020
Streptomyces mobaraensis (P81453)
Fotschki, J.; Wroblewska, B.; Fotschki, B.; Kalicki, B.; Rigby, N.; Mackie, A.
Microbial transglutaminase alters the immunogenic potential and cross-reactivity of horse and cow milk proteins
J. Dairy Sci.
103
2153-2166
2020
Streptomyces mobaraensis (P81453)
Jin, M.; Huang, J.; Pei, Z.; Huang, J.; Gao, H.; Chang, Z.
Purification and characterization of a high-salt-resistant microbial transglutaminase from Streptomyces mobaraensis
J. Mol. Catal. B
133
6-11
2016
Streptomyces mobaraensis (P81453), Streptomyces mobaraensis TX (P81453)
-
Long, H.; Ma, K.; Xiao, Z.; Ren, X.; Yang, G.
Preparation and characteristics of gelatin sponges crosslinked by microbial transglutaminase
PeerJ
5
e3665
2017
Streptomyces mobaraensis (P81453)
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