Application | Comment | Organism |
---|---|---|
synthesis | structure-based modification of D-alanine-D-alanine ligase from strain ATCC 43589 for D-alanyl-D-lactate and other depsipeptide synthesis by mutant S137G/Y207F | Thermotoga maritima |
Cloned (Comment) | Organism |
---|---|
expression of mutant enzymes in Escherichia coli strain BL21(DE3) as His6-tagged proteins | Thermotoga maritima |
Protein Variants | Comment | Organism |
---|---|---|
additional information | structure-based modification of D-alanine-D-alanine ligase from strain ATCC 43589 for depsipeptide synthesis, overview | Thermotoga maritima |
S137A | site-directed mutagenesis, the mutant D-alanine-D-alanine ligase also shows formation of D-alanyl-D-lactate depsipeptide in contrast to the wild-type enzyme, EC 6.3.2.4, which is inactive with (R)-lactate | Thermotoga maritima |
S137A/Y207F | site-directed mutagenesis, the mutant D-alanine-D-alanine ligase also shows formation of D-alanyl-D-lactate depsipeptide in contrast to the wild-type enzyme, EC 6.3.2.4, which is inactive with (R)-lactate | Thermotoga maritima |
S137F/Y207F | site-directed mutagenesis, the mutant D-alanine-D-alanine ligase also shows formation of D-alanyl-D-lactate depsipeptide in contrast to the wild-type enzyme, EC 6.3.2.4, which is inactive with (R)-lactate | Thermotoga maritima |
S137G/Y207F | site-directed mutagenesis, the mutant D-alanine-D-alanine ligase also shows formation of D-alanyl-D-lactate depsipeptide in contrast to the wild-type enzyme, EC 6.3.2.4, which is inactive with (R)-lactate | Thermotoga maritima |
S137T/Y207F | site-directed mutagenesis, the mutant D-alanine-D-alanine ligase also shows formation of D-alanyl-D-lactate depsipeptide in contrast to the wild-type enzyme, EC 6.3.2.4, which is inactive with (R)-lactate | Thermotoga maritima |
Y201F/Y207F | site-directed mutagenesis, the mutant D-alanine-D-alanine ligase also shows formation of D-alanyl-D-lactate depsipeptide in contrast to the wild-type enzyme, EC 6.3.2.4, which is inactive with (R)-lactate | Thermotoga maritima |
Y207F | site-directed mutagenesis, the mutant D-alanine-D-alanine ligase also shows formation of D-alanyl-D-lactate depsipeptide in contrast to the wild-type enzyme, EC 6.3.2.4, which is inactive with (R)-lactate | Thermotoga maritima |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermotoga maritima | - |
- |
- |
Thermotoga maritima ATCC 43589 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Thermotoga maritima |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.19 | - |
D-alanyl-D-lactate depsipeptide formation activity, recombinant D-alanine-D-alanine ligase mutant S137F/Y207F, pH 7.5, 60°C | Thermotoga maritima |
0.39 | - |
D-alanyl-D-lactate depsipeptide formation activity, recombinant D-alanine-D-alanine ligase mutant Y207F, pH 7.5, 60°C | Thermotoga maritima |
0.42 | - |
D-alanyl-D-lactate depsipeptide formation activity, recombinant D-alanine-D-alanine ligase mutant S137A/Y207F, pH 7.5, 60°C | Thermotoga maritima |
1.2 | - |
D-alanyl-D-lactate depsipeptide formation activity, recombinant D-alanine-D-alanine ligase mutant S137G/Y207F, pH 7.5, 60°C | Thermotoga maritima |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-alanine + (R)-lactate + ATP | mutants variants Y207F, S137G/Y207F, S137F/Y207F, S137T/Y207F, and S137A/Y207F from D-alanine-D-alanine ligase, EC 6.3.2.4. The wild-type D-alanine-D-alanine ligase, EC 6.3.2.4 does not show activity with (R)-lactate | Thermotoga maritima | D-alanyl-(R)-lactate + ADP + phosphate | - |
? | |
D-alanine + (R)-lactate + ATP | mutants variants Y207F, S137G/Y207F, S137F/Y207F, S137T/Y207F, and S137A/Y207F from D-alanine-D-alanine ligase, EC 6.3.2.4. The wild-type D-alanine-D-alanine ligase, EC 6.3.2.4 does not show activity with (R)-lactate | Thermotoga maritima ATCC 43589 | D-alanyl-(R)-lactate + ADP + phosphate | - |
? | |
additional information | substrate specificity of recombinant D-alanine-D-alanine ligase mutant S137G/Y207F, overview | Thermotoga maritima | ? | - |
? | |
additional information | substrate specificity of recombinant D-alanine-D-alanine ligase mutant S137G/Y207F, overview | Thermotoga maritima ATCC 43589 | ? | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
assay at | Thermotoga maritima |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Thermotoga maritima |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Thermotoga maritima |