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Literature summary extracted from

  • Mu, D.; Li, H.; Chen, Q.; Zhu, J.; Wu, X.; Luo, S.; Zhao, Y.; Wang, L.; Jiang, S.; Li, X.; Zheng, Z.
    Secretion of Bacillus amyloliquefaciens gamma-glutamyltranspeptidase from Bacillus subtilis and its application in enzymatic synthesis of L-theanine (2019), J. Agric. Food Chem., 67, 14129-14136 .
    View publication on PubMed

Application

EC Number Application Comment Organism
2.3.2.2 synthesis the recombinant extracellular enzyme is used for enzymatic synthesis of L-theanine, which is performed at optimal conditions: 20 mM L-Gln, 100 mM ethylamine-HCl, 0.5 U/ml BaGGT, incubated at 40°C for 6 h, 200 rpm Bacillus amyloliquefaciens

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.2.2 gene ggt, recombinant expression of His6-tagged enzyme in Bacillus subtilis strain 168 (pHT43-spAmyQ-ggtDELTAsp) under control of the Plac promoter, extracellular secretion, subcloning in Escherichia coli strain DH5alpha, method optimization. Dual-signal peptides promote more secretion than a single signal peptide Bacillus amyloliquefaciens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.2.2 0.214
-
 L-gamma-glutamyl-4-nitroanilide recombinant extracellular enzyme, pH 9.0, 40°C Bacillus amyloliquefaciens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.2.2 a (5-L-glutamyl)-peptide + an amino acid Bacillus amyloliquefaciens
-
 a peptide + a 5-L-glutamyl amino acid
-
 ?
2.3.2.2 a (5-L-glutamyl)-peptide + an amino acid Bacillus amyloliquefaciens BH072
-
 a peptide + a 5-L-glutamyl amino acid
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.2.2 Bacillus amyloliquefaciens
-
-
-
2.3.2.2 Bacillus amyloliquefaciens BH072
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.2.2 recombinant His6-tagged eextracellular enzyme from Bacillus subtilis strain 168 by nickel affinity chromatography Bacillus amyloliquefaciens

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.3.2.2 55
-
 purified recombinant extracellular enzyme, pH 9.0, 40°C, substrate L-gamma-glutamyl-4-nitroanilide Bacillus amyloliquefaciens

Storage Stability

EC Number Storage Stability Organism
2.3.2.2 40°C, purified recombinant extracellular enzyme, 72 h, 90% of initial activity remaining, and more than 50% of initial activity after being maintained at different pH values for 24 h Bacillus amyloliquefaciens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.2.2 a (5-L-glutamyl)-peptide + an amino acid
-
 Bacillus amyloliquefaciens a peptide + a 5-L-glutamyl amino acid
-
 ?
2.3.2.2 a (5-L-glutamyl)-peptide + an amino acid
-
 Bacillus amyloliquefaciens BH072 a peptide + a 5-L-glutamyl amino acid
-
 ?
2.3.2.2 L-gamma-glutamyl-4-nitroanilide + glycylglycine
-
 Bacillus amyloliquefaciens 4-nitroaniline + 5-L-glutamyl-glycylglycine
-
 ?
2.3.2.2 L-gamma-glutamyl-4-nitroanilide + glycylglycine
-
 Bacillus amyloliquefaciens BH072 4-nitroaniline + 5-L-glutamyl-glycylglycine
-
 ?
2.3.2.2 L-glutamine + ethylamine GGT transfers the gamma-glutamyl moiety from L-Gln to ethylamine to form the product gamma-glutamylethylamine (L-theanine), product identification by mass spectrometry, optimization of reaction conditions, overview Bacillus amyloliquefaciens NH3 + gamma-glutamylethylamine
-
 ?
2.3.2.2 L-glutamine + ethylamine GGT transfers the gamma-glutamyl moiety from L-Gln to ethylamine to form the product gamma-glutamylethylamine (L-theanine), product identification by mass spectrometry, optimization of reaction conditions, overview Bacillus amyloliquefaciens BH072 NH3 + gamma-glutamylethylamine
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.2.2 heterodimer 1 * 46000, large subunit, + 1 * 22000, small subunit, His6-tagged enzyme, SDS-PAGE Bacillus amyloliquefaciens

Synonyms

EC Number Synonyms Comment Organism
2.3.2.2 BaGGT
-
 Bacillus amyloliquefaciens
2.3.2.2 gamma-glutamyltranspeptidase
-
 Bacillus amyloliquefaciens
2.3.2.2 GGT
-
 Bacillus amyloliquefaciens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.2.2 40
-
 recombinant enzyme Bacillus amyloliquefaciens

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.3.2.2 20 70 40/56/34/20/6% relative activity at 20/30/50/60/70°C, respectively, recombinant enzyme, profile overview Bacillus amyloliquefaciens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.3.2.2 40 50 there is hardly any activity loss of the purified recombinant enzyme after preincubation at 40 °C for 72 h, but the activity drops to 7.5% after preincubation for 8 h at 50°C Bacillus amyloliquefaciens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.2.2 9
-
 recombinant enzyme Bacillus amyloliquefaciens

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.3.2.2 5 11 46/75/64% relative activity at pH 7.0/8.0/10.0, respectively, recombinant enzyme, profile overview Bacillus amyloliquefaciens

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.3.2.2 5 11 25°C, 7 h, 80% activity or higher is retained at pH 7.0-10.0 and, in all cases, it is above 50% at pH 5.0-11.0 Bacillus amyloliquefaciens