Protein Variants | Comment | Organism |
---|---|---|
additional information | easy and efficient method for GGT enzyme immobilization, preparation of thermo- and salt-tolerant chitosan cross-linked gamma-glutamyl transpeptidase from Bacillus licheniformis strain ER15, overview. The purified native enzyme is covalently immobilized onto chitosan microspheres (CMS) standardized with respect to pH, enzyme load, and time. Immobilization efficiency of 11.9 U/mg dry weight of microsphere is obtained in Tris-HCl, pH 9.0, at 18°C in 4 h. Immobilized enzyme CMS-GGT exhibits improved thermal stability (t1/2 of 70.7 min at 60°C), activity in a broader pH range and improved salt stability in 18% (3 M) sodium chloride solution as compared to free enzyme. Both free and immobilized enzymes specifically convert glutamine to glutamic acid in a mixture of amino acids. CMS-GGT has a better shelf life and high recyclability retaining 90% catalytic efficiency up to 10 reaction cycles compared to free enzyme. For long-term storage, CMS-GGT can be disinfected using either sodium azide or sodium hypochlorite solution without affecting the enzyme activity | Bacillus licheniformis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1,10-phenanthroline | slight inhibition of free and immobilized enzyme | Bacillus licheniformis | |
2-mercaptoethanol | slight inhibition of free and immobilized enzyme | Bacillus licheniformis | |
6-diazo-5-oxo-L-norleucine | i.e. DON | Bacillus licheniformis | |
azaserine | complete inhibition of the free enzyme | Bacillus licheniformis | |
Bromoacetic acid | slight inhibition of free enzyme | Bacillus licheniformis | |
Ca2+ | inhibits the free enzyme by about 50% at 5 mM | Bacillus licheniformis | |
Cd2+ | inhibits the free enzyme by about 75% and the immobilized enzyme by about 20% at 5 mM | Bacillus licheniformis | |
Co2+ | inhibits the free enzyme by about 70% and the immobilized enzyme by about 15% at 5 mM | Bacillus licheniformis | |
Cu2+ | inhibits the free enzyme by about 50% at 5 mM | Bacillus licheniformis | |
DTT | slight inhibition of free and immobilized enzyme | Bacillus licheniformis | |
EDTA | - |
Bacillus licheniformis | |
EGTA | - |
Bacillus licheniformis | |
iodoacetamide | - |
Bacillus licheniformis | |
iodoacetic acid | slight inhibition of free and immobilized enzyme | Bacillus licheniformis | |
Mg2+ | inhibits the free enzyme by about 15% at 5 mM | Bacillus licheniformis | |
Mn2+ | inhibits the free enzyme by about 50% at 5 mM | Bacillus licheniformis | |
additional information | immobilized enzyme CMS-GGT is 10-20% less susceptibile to inhibition by PMSF, N-bromosuccinamide and GGT specific inhibitors azaserine and 6-diazo-5-oxo-L-norleucine as compared to free enzyme | Bacillus licheniformis | |
N-bromosuccinamide | - |
Bacillus licheniformis | |
NaCl | about 45% inhibition of the free enzyme at 2-18% w/v NaCl, about 20% inhibition of the immobilized enzyme | Bacillus licheniformis | |
NaN3 | slight inhibition of free and immobilized enzyme | Bacillus licheniformis | |
Ni2+ | inhibits the free enzyme by about 25% and the immobilized enzyme by about 20% at 5 mM | Bacillus licheniformis | |
Pb2+ | inhibits the free enzyme by about 65% and the immobilized enzyme by about 25% at 5 mM | Bacillus licheniformis | |
PMSF | - |
Bacillus licheniformis | |
Zn2+ | inhibits the free enzyme by about 85% and the immobilized enzyme by about 5% at 5 mM | Bacillus licheniformis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics of purified free enzyme BLGGT and purified immobilized enzyme CMS-GGT | Bacillus licheniformis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | activates the immobilized enzyme by about 20% at 5 mM | Bacillus licheniformis | |
Cu2+ | activates the immobilized enzyme by about 20% at 5 mM | Bacillus licheniformis | |
Mg2+ | activates the free enzyme by about 5% at 5 mM | Bacillus licheniformis | |
additional information | Mn2+ has no effect on the immobilized enzyme at 5 mM | Bacillus licheniformis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
a (5-L-glutamyl)-peptide + an amino acid | Bacillus licheniformis | - |
a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | Bacillus licheniformis ER15 | - |
a peptide + a 5-L-glutamyl amino acid | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus licheniformis | A9YTT0 | - |
- |
Bacillus licheniformis ER15 | A9YTT0 | - |
- |
Purification (Comment) | Organism |
---|---|
native extracellular enzyme 4.6fold from Bacillus licheniformis strain ER15 by ultrafiltration, gel filtration, ion exchange chromatography, and dialysis, with 50.11% yield | Bacillus licheniformis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
250.09 | - |
purified free enzyme, pH and temperature not specified in the publication | Bacillus licheniformis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
a (5-L-glutamyl)-peptide + an amino acid | - |
Bacillus licheniformis | a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | - |
Bacillus licheniformis ER15 | a peptide + a 5-L-glutamyl amino acid | - |
? | |
L-gamma-glutamyl-4-nitroanilide + ethylamine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-ethylamine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + ethylamine | - |
Bacillus licheniformis ER15 | 4-nitroaniline + 5-L-glutamyl-ethylamine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-glycine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycine | - |
Bacillus licheniformis ER15 | 4-nitroaniline + 5-L-glutamyl-glycine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycylglycine | best substrate | Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycylglycine | best substrate | Bacillus licheniformis ER15 | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-alanine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-alanine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-arginine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-arginine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-asparagine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-asparagine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-aspartate | low activity | Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-aspartate | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamate | low activity | Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-glutamate | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-glutamine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-histidine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-histidine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-isoleucine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-isoleucine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-leucine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-leucine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-lysine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-lysine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-methionine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-methionine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-phenylalanine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-phenylalanine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-serine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-serine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-threonine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-threonine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-tryptophan | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-tryptophan | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-valine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-L-valine | - |
? | |
additional information | both free and immobilized enzymes specifically convert glutamine to glutamic acid in a mixture of amino acids | Bacillus licheniformis | ? | - |
- |
|
additional information | both free and immobilized enzymes specifically convert glutamine to glutamic acid in a mixture of amino acids | Bacillus licheniformis ER15 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
BlGGT | - |
Bacillus licheniformis |
gamma-glutamyl transpeptidase | - |
Bacillus licheniformis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
free and immobilized enzymes | Bacillus licheniformis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 80 | about 60% of maximal activity at 40°C for free and immobilized enzyme, 30% of maximal activity at 80°C for the free enzyme, 75% for the immbilized enzyme, profile overview | Bacillus licheniformis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
free and immobilized enzymes | Bacillus licheniformis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 11 | over 40% of maximal activity within this range for the free enzyme, profile overview | Bacillus licheniformis |
5 | 12 | over 60% of maximal activity within this range for the immobilized enzyme, profile overview | Bacillus licheniformis |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5 | 12 | free and immobilized enzymes, stable at with 90-100% activity remaining | Bacillus licheniformis |