BRENDA - Enzyme Database

Overproduction of the Bacillus subtilis glutamyl-tRNA synthetase in its host and its toxicity to Escherichia coli

Pelchat, M.; Lacoste, L.; Yang, F.; Lapointe, J.; Can. J. Microbiol. 44, 378-381 (1998)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
6.1.1.17
gene gltX, expression in Escherichia coli is lethal, but a modulated enzyme, lacking a part of the tRNA acceptor stem binding domain, can be recombinantly expressed in Escherichia coli DH5alpha, overexpression in Bacillus subtilis
Bacillus subtilis
6.1.1.24
the gene is cloned with its sigmaA promoter and a downstream region including a rho-independent terminator in the shuttle vector pRB394 for Escherichia coli and bacillus subtilis. Transformation of Bacillus subtilis with this recombinant plasmidleads to a 30fold increase of glutamyl-tRNA synthetase specific activity in crude extracts. Transformation of Escherichia coli with this plasmid gives no recombinants. The presence of Bacillus subtilis glutamyl-tRNA synthetase is lethal for Escherichia coli, probably because this enzyme glutamylates tRNA1Gln in vivo as it does in vitro
Bacillus subtilis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
6.1.1.17
55000
-
x * 55000, recombinant enzyme expressed in Bacillus subtilis, SDS-PAGE
Bacillus subtilis
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
6.1.1.17
ATP + L-glutamate + tRNAGln
Bacillus subtilis
-
AMP + diphosphate + L-glutamyl-tRNAGln
-
Bacillus subtilis
?
6.1.1.17
ATP + L-glutamate + tRNAGlu
Bacillus subtilis
-
AMP + diphosphate + L-glutamyl-tRNAGlu
-
Bacillus subtilis
?
6.1.1.17
additional information
Bacillus subtilis
the recombinant wild-type enzyme is toxic for Escherichia coli, probably due to its charging of both tRNAGlu and tRNAGln
?
-
Bacillus subtilis
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
6.1.1.17
Bacillus subtilis
-
-
-
6.1.1.24
Bacillus subtilis
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
6.1.1.17
50fold, recombinant enzyme from overexpression in Bacillus subtilis, to homogeneity
Bacillus subtilis
6.1.1.24
-
Bacillus subtilis
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
6.1.1.17
0.16
-
purified recombinant enzyme expressed in Bacillus subtilis
Bacillus subtilis
6.1.1.24
0.16
-
-
Bacillus subtilis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.1.1.17
ATP + L-glutamate + tRNAGln
-
492295
Bacillus subtilis
AMP + diphosphate + L-glutamyl-tRNAGln
-
492295
Bacillus subtilis
?
6.1.1.17
ATP + L-glutamate + tRNAGlu
-
492295
Bacillus subtilis
AMP + diphosphate + L-glutamyl-tRNAGlu
-
492295
Bacillus subtilis
?
6.1.1.17
additional information
the recombinant wild-type enzyme is toxic for Escherichia coli, probably due to its charging of both tRNAGlu and tRNAGln
492295
Bacillus subtilis
?
-
492295
Bacillus subtilis
?
6.1.1.24
ATP + L-glutamate + tRNAGln
-
492295
Bacillus subtilis
AMP + diphosphate + Glu-tRNAGln
-
-
-
?
6.1.1.24
ATP + L-glutamate + tRNAGlu
-
492295
Bacillus subtilis
AMP + diphosphate + Glu-tRNAGlu
-
492295
Bacillus subtilis
?
Subunits
EC Number
Subunits
Commentary
Organism
6.1.1.17
?
x * 55000, recombinant enzyme expressed in Bacillus subtilis, SDS-PAGE
Bacillus subtilis
Temperature Optimum [░C]
EC Number
Temperature Optimum [░C]
Temperature Optimum Maximum [░C]
Commentary
Organism
6.1.1.17
37
-
assay at
Bacillus subtilis
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
6.1.1.17
ATP
-
Bacillus subtilis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
6.1.1.17
gene gltX, expression in Escherichia coli is lethal, but a modulated enzyme, lacking a part of the tRNA acceptor stem binding domain, can be recombinantly expressed in Escherichia coli DH5alpha, overexpression in Bacillus subtilis
Bacillus subtilis
6.1.1.24
the gene is cloned with its sigmaA promoter and a downstream region including a rho-independent terminator in the shuttle vector pRB394 for Escherichia coli and bacillus subtilis. Transformation of Bacillus subtilis with this recombinant plasmidleads to a 30fold increase of glutamyl-tRNA synthetase specific activity in crude extracts. Transformation of Escherichia coli with this plasmid gives no recombinants. The presence of Bacillus subtilis glutamyl-tRNA synthetase is lethal for Escherichia coli, probably because this enzyme glutamylates tRNA1Gln in vivo as it does in vitro
Bacillus subtilis
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
6.1.1.17
ATP
-
Bacillus subtilis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
6.1.1.17
55000
-
x * 55000, recombinant enzyme expressed in Bacillus subtilis, SDS-PAGE
Bacillus subtilis
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
6.1.1.17
ATP + L-glutamate + tRNAGln
Bacillus subtilis
-
AMP + diphosphate + L-glutamyl-tRNAGln
-
Bacillus subtilis
?
6.1.1.17
ATP + L-glutamate + tRNAGlu
Bacillus subtilis
-
AMP + diphosphate + L-glutamyl-tRNAGlu
-
Bacillus subtilis
?
6.1.1.17
additional information
Bacillus subtilis
the recombinant wild-type enzyme is toxic for Escherichia coli, probably due to its charging of both tRNAGlu and tRNAGln
?
-
Bacillus subtilis
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
6.1.1.17
50fold, recombinant enzyme from overexpression in Bacillus subtilis, to homogeneity
Bacillus subtilis
6.1.1.24
-
Bacillus subtilis
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
6.1.1.17
0.16
-
purified recombinant enzyme expressed in Bacillus subtilis
Bacillus subtilis
6.1.1.24
0.16
-
-
Bacillus subtilis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.1.1.17
ATP + L-glutamate + tRNAGln
-
492295
Bacillus subtilis
AMP + diphosphate + L-glutamyl-tRNAGln
-
492295
Bacillus subtilis
?
6.1.1.17
ATP + L-glutamate + tRNAGlu
-
492295
Bacillus subtilis
AMP + diphosphate + L-glutamyl-tRNAGlu
-
492295
Bacillus subtilis
?
6.1.1.17
additional information
the recombinant wild-type enzyme is toxic for Escherichia coli, probably due to its charging of both tRNAGlu and tRNAGln
492295
Bacillus subtilis
?
-
492295
Bacillus subtilis
?
6.1.1.24
ATP + L-glutamate + tRNAGln
-
492295
Bacillus subtilis
AMP + diphosphate + Glu-tRNAGln
-
-
-
?
6.1.1.24
ATP + L-glutamate + tRNAGlu
-
492295
Bacillus subtilis
AMP + diphosphate + Glu-tRNAGlu
-
492295
Bacillus subtilis
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
6.1.1.17
?
x * 55000, recombinant enzyme expressed in Bacillus subtilis, SDS-PAGE
Bacillus subtilis
Temperature Optimum [░C] (protein specific)
EC Number
Temperature Optimum [░C]
Temperature Optimum Maximum [░C]
Commentary
Organism
6.1.1.17
37
-
assay at
Bacillus subtilis