Cloned (Comment) | Organism |
---|---|
gene plsXX, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain C43(DE3) | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
E236A | site-directed mutagenesis | Bacillus subtilis |
K184A | site-directed mutagenesis | Bacillus subtilis |
K184Q | site-directed mutagenesis | Bacillus subtilis |
K184R | site-directed mutagenesis | Bacillus subtilis |
K233A | site-directed mutagenesis | Bacillus subtilis |
N229A | site-directed mutagenesis, the mutation causes the greatest kcat decrease by more than 10fold and increases KM by 4.4 and 2.6fold for palmitoyl-ACP and phosphate, respectively | Bacillus subtilis |
N229Q | site-directed mutagenesis, most of the catalytic activity of mutant N229A is recovered with a 2.2fold KM increase for palmitoyl-ACP when glutamine is introduced into the mutated position in N229Q | Bacillus subtilis |
R114A | site-directed mutagenesis, the catalytic activity of the negative control mutant protein R114A is essentially the same compared to wild-type enzyme for both substrates | Bacillus subtilis |
R120A | site-directed mutagenesis, the mutation causes a significant 3.3fold increase in KM for palmitoyl-ACP without adverse effect on other kinetic constants | Bacillus subtilis |
R73A | site-directed mutagenesis, the mutation causes a significant 2.5fold increase in KM for palmitoyl-ACP without adverse effect on other kinetic constants | Bacillus subtilis |
R73A/R120A | site-directed mutagenesis, the double mutation diminishes the enzyme activity to an undetectable level | Bacillus subtilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics of enzyme PlsX | Bacillus subtilis | |
0.00039 | - |
holo-[acyl-carrier protein] | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
0.00108 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant E236A | Bacillus subtilis | |
0.00124 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant E209A | Bacillus subtilis | |
0.00135 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant E181A | Bacillus subtilis | |
0.00139 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant K184R | Bacillus subtilis | |
0.0014 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
0.00143 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant K233A | Bacillus subtilis | |
0.00149 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant K184A | Bacillus subtilis | |
0.0017 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant R114A | Bacillus subtilis | |
0.00202 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant K184Q | Bacillus subtilis | |
0.00304 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant N229Q | Bacillus subtilis | |
0.00336 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant R120A | Bacillus subtilis | |
0.0037 | - |
palmitoyl phosphate | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
0.0045 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant R73A | Bacillus subtilis | |
0.006 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant N229A | Bacillus subtilis | |
0.052 | - |
phosphate | pH 7.5, 37°C, mutant K184R | Bacillus subtilis | |
0.093 | - |
phosphate | pH 7.5, 37°C, mutant E209A | Bacillus subtilis | |
0.104 | - |
phosphate | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
0.111 | - |
phosphate | pH 7.5, 37°C, mutant R73A | Bacillus subtilis | |
0.126 | - |
phosphate | pH 7.5, 37°C, mutant N229Q | Bacillus subtilis | |
0.127 | - |
phosphate | pH 7.5, 37°C, mutant R114A | Bacillus subtilis | |
0.131 | - |
phosphate | pH 7.5, 37°C, mutant R120A | Bacillus subtilis | |
0.136 | - |
phosphate | pH 7.5, 37°C, mutant K233A | Bacillus subtilis | |
0.151 | - |
phosphate | pH 7.5, 37°C, mutant E236A | Bacillus subtilis | |
0.239 | - |
phosphate | pH 7.5, 37°C, mutant E181A | Bacillus subtilis | |
0.267 | - |
phosphate | pH 7.5, 37°C, mutant N229A | Bacillus subtilis | |
0.425 | - |
phosphate | pH 7.5, 37°C, mutant K184Q | Bacillus subtilis | |
0.802 | - |
phosphate | pH 7.5, 37°C, mutant K184A | Bacillus subtilis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
palmitoyl-[acyl-carrier protein] + phosphate | Bacillus subtilis | - |
palmitoyl phosphate + holo-[acyl-carrier protein] | - |
r | |
palmitoyl-[acyl-carrier protein] + phosphate | Bacillus subtilis 168 | - |
palmitoyl phosphate + holo-[acyl-carrier protein] | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | P71018 | - |
- |
Bacillus subtilis 168 | P71018 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain C43(DE3) | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
an acyl-[acyl-carrier protein] + phosphate | - |
Bacillus subtilis | an acyl phosphate + an [acyl-carrier protein] | - |
r | |
an acyl-[acyl-carrier protein] + phosphate | - |
Bacillus subtilis 168 | an acyl phosphate + an [acyl-carrier protein] | - |
r | |
palmitoyl-[acyl-carrier protein] + phosphate | - |
Bacillus subtilis | palmitoyl phosphate + holo-[acyl-carrier protein] | - |
r | |
palmitoyl-[acyl-carrier protein] + phosphate | the reverse reaction is highly preferred | Bacillus subtilis | palmitoyl phosphate + holo-[acyl-carrier protein] | - |
r | |
palmitoyl-[acyl-carrier protein] + phosphate | - |
Bacillus subtilis 168 | palmitoyl phosphate + holo-[acyl-carrier protein] | - |
r | |
palmitoyl-[acyl-carrier protein] + phosphate | the reverse reaction is highly preferred | Bacillus subtilis 168 | palmitoyl phosphate + holo-[acyl-carrier protein] | - |
r |
Synonyms | Comment | Organism |
---|---|---|
phosphate: acyl-ACP acyltransferase | - |
Bacillus subtilis |
phosphate: acyl-acyl carrier protein acyltransferase | - |
Bacillus subtilis |
plsX | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Bacillus subtilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001 | - |
phosphate | pH 7.5, 37°C, mutant N229A | Bacillus subtilis | |
0.001 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant N229A | Bacillus subtilis | |
0.00127 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
0.002 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant K184A | Bacillus subtilis | |
0.0023 | - |
phosphate | pH 7.5, 37°C, mutant K184A | Bacillus subtilis | |
0.0025 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant K184Q | Bacillus subtilis | |
0.0028 | - |
phosphate | pH 7.5, 37°C, mutant K184Q | Bacillus subtilis | |
0.003 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant K233A | Bacillus subtilis | |
0.0033 | - |
phosphate | pH 7.5, 37°C, mutant E209A | Bacillus subtilis | |
0.0033 | - |
phosphate | pH 7.5, 37°C, mutant K233A | Bacillus subtilis | |
0.0036 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant E209A | Bacillus subtilis | |
0.0042 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant E236A | Bacillus subtilis | |
0.0043 | - |
phosphate | pH 7.5, 37°C, mutant E181A | Bacillus subtilis | |
0.0045 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant E181A | Bacillus subtilis | |
0.0045 | - |
phosphate | pH 7.5, 37°C, mutant E236A | Bacillus subtilis | |
0.0067 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant K184R | Bacillus subtilis | |
0.0068 | - |
phosphate | pH 7.5, 37°C, mutant K184R | Bacillus subtilis | |
0.0103 | - |
phosphate | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
0.012 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant R114A | Bacillus subtilis | |
0.012 | - |
phosphate | pH 7.5, 37°C, mutant R73A | Bacillus subtilis | |
0.0125 | - |
phosphate | pH 7.5, 37°C, mutant N229Q | Bacillus subtilis | |
0.0142 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant R73A | Bacillus subtilis | |
0.0145 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant N229Q | Bacillus subtilis | |
0.122 | - |
phosphate | pH 7.5, 37°C, mutant R114A | Bacillus subtilis | |
0.123 | - |
phosphate | pH 7.5, 37°C, mutant R120A | Bacillus subtilis | |
0.128 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, mutant R120A | Bacillus subtilis | |
2.467 | - |
holo-[acyl-carrier protein] | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
2.783 | - |
palmitoyl phosphate | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Bacillus subtilis |
General Information | Comment | Organism |
---|---|---|
additional information | substrate recognition and catalytic mechanism of phosphate acyltransferase PlsX, docking simulation, detailed overview. Conservation of potential active-site residues, sequence comparisons. The suspected ACP docking site is formed by Arg73 and Arg120 | Bacillus subtilis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.099 | - |
phosphate | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
0.907 | - |
palmitoyl-[acyl-carrier protein] | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
752.3 | - |
palmitoyl phosphate | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
6325.6 | - |
holo-[acyl-carrier protein] | pH 7.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis |