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Literature summary extracted from

  • Jiang, Y.; Qin, M.; Guo, Z.
    Substrate recognition and catalytic mechanism of the phosphate acyltransferase PlsX from Bacillus subtilis (2020), ChemBioChem, 21, 1-11 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.274 gene plsXX, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain C43(DE3) Bacillus subtilis

Protein Variants

EC Number Protein Variants Comment Organism
2.3.1.274 E236A site-directed mutagenesis Bacillus subtilis
2.3.1.274 K184A site-directed mutagenesis Bacillus subtilis
2.3.1.274 K184Q site-directed mutagenesis Bacillus subtilis
2.3.1.274 K184R site-directed mutagenesis Bacillus subtilis
2.3.1.274 K233A site-directed mutagenesis Bacillus subtilis
2.3.1.274 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
2.3.1.274 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
2.3.1.274 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
2.3.1.274 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
2.3.1.274 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
2.3.1.274 R73A/R120A site-directed mutagenesis, the double mutation diminishes the enzyme activity to an undetectable level Bacillus subtilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.274 additional information
-
 additional information steady-state kinetics of enzyme PlsX Bacillus subtilis
2.3.1.274 0.00039
-
 holo-[acyl-carrier protein] pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis
2.3.1.274 0.00108
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant E236A Bacillus subtilis
2.3.1.274 0.00124
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant E209A Bacillus subtilis
2.3.1.274 0.00135
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant E181A Bacillus subtilis
2.3.1.274 0.00139
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant K184R Bacillus subtilis
2.3.1.274 0.0014
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis
2.3.1.274 0.00143
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant K233A Bacillus subtilis
2.3.1.274 0.00149
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant K184A Bacillus subtilis
2.3.1.274 0.0017
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant R114A Bacillus subtilis
2.3.1.274 0.00202
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant K184Q Bacillus subtilis
2.3.1.274 0.00304
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant N229Q Bacillus subtilis
2.3.1.274 0.00336
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant R120A Bacillus subtilis
2.3.1.274 0.0037
-
 palmitoyl phosphate pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis
2.3.1.274 0.0045
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant R73A Bacillus subtilis
2.3.1.274 0.006
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant N229A Bacillus subtilis
2.3.1.274 0.052
-
 phosphate pH 7.5, 37°C, mutant K184R Bacillus subtilis
2.3.1.274 0.093
-
 phosphate pH 7.5, 37°C, mutant E209A Bacillus subtilis
2.3.1.274 0.104
-
 phosphate pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis
2.3.1.274 0.111
-
 phosphate pH 7.5, 37°C, mutant R73A Bacillus subtilis
2.3.1.274 0.126
-
 phosphate pH 7.5, 37°C, mutant N229Q Bacillus subtilis
2.3.1.274 0.127
-
 phosphate pH 7.5, 37°C, mutant R114A Bacillus subtilis
2.3.1.274 0.131
-
 phosphate pH 7.5, 37°C, mutant R120A Bacillus subtilis
2.3.1.274 0.136
-
 phosphate pH 7.5, 37°C, mutant K233A Bacillus subtilis
2.3.1.274 0.151
-
 phosphate pH 7.5, 37°C, mutant E236A Bacillus subtilis
2.3.1.274 0.239
-
 phosphate pH 7.5, 37°C, mutant E181A Bacillus subtilis
2.3.1.274 0.267
-
 phosphate pH 7.5, 37°C, mutant N229A Bacillus subtilis
2.3.1.274 0.425
-
 phosphate pH 7.5, 37°C, mutant K184Q Bacillus subtilis
2.3.1.274 0.802
-
 phosphate pH 7.5, 37°C, mutant K184A Bacillus subtilis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.1.274 palmitoyl-[acyl-carrier protein] + phosphate Bacillus subtilis
-
 palmitoyl phosphate + holo-[acyl-carrier protein]
-
 r
2.3.1.274 palmitoyl-[acyl-carrier protein] + phosphate Bacillus subtilis 168
-
 palmitoyl phosphate + holo-[acyl-carrier protein]
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.274 Bacillus subtilis P71018
-
-
2.3.1.274 Bacillus subtilis 168 P71018
-
-
2.3.1.276 Bacillus subtilis
-
-
-
2.3.1.276 Bacillus subtilis 168
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.274 recombinant wild-type and mutant enzymes from Escherichia coli strain C43(DE3) Bacillus subtilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.274 an acyl-[acyl-carrier protein] + phosphate
-
 Bacillus subtilis an acyl phosphate + an [acyl-carrier protein]
-
 r
2.3.1.274 an acyl-[acyl-carrier protein] + phosphate
-
 Bacillus subtilis 168 an acyl phosphate + an [acyl-carrier protein]
-
 r
2.3.1.274 palmitoyl-[acyl-carrier protein] + phosphate
-
 Bacillus subtilis palmitoyl phosphate + holo-[acyl-carrier protein]
-
 r
2.3.1.274 palmitoyl-[acyl-carrier protein] + phosphate the reverse reaction is highly preferred Bacillus subtilis palmitoyl phosphate + holo-[acyl-carrier protein]
-
 r
2.3.1.274 palmitoyl-[acyl-carrier protein] + phosphate
-
 Bacillus subtilis 168 palmitoyl phosphate + holo-[acyl-carrier protein]
-
 r
2.3.1.274 palmitoyl-[acyl-carrier protein] + phosphate the reverse reaction is highly preferred Bacillus subtilis 168 palmitoyl phosphate + holo-[acyl-carrier protein]
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.3.1.274 phosphate: acyl-ACP acyltransferase
-
 Bacillus subtilis
2.3.1.274 phosphate: acyl-acyl carrier protein acyltransferase
-
 Bacillus subtilis
2.3.1.274 plsX
-
 Bacillus subtilis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.1.274 37
-
 assay at Bacillus subtilis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.1.274 0.001
-
 phosphate pH 7.5, 37°C, mutant N229A Bacillus subtilis
2.3.1.274 0.001
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant N229A Bacillus subtilis
2.3.1.274 0.00127
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis
2.3.1.274 0.002
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant K184A Bacillus subtilis
2.3.1.274 0.0023
-
 phosphate pH 7.5, 37°C, mutant K184A Bacillus subtilis
2.3.1.274 0.0025
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant K184Q Bacillus subtilis
2.3.1.274 0.0028
-
 phosphate pH 7.5, 37°C, mutant K184Q Bacillus subtilis
2.3.1.274 0.003
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant K233A Bacillus subtilis
2.3.1.274 0.0033
-
 phosphate pH 7.5, 37°C, mutant E209A Bacillus subtilis
2.3.1.274 0.0033
-
 phosphate pH 7.5, 37°C, mutant K233A Bacillus subtilis
2.3.1.274 0.0036
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant E209A Bacillus subtilis
2.3.1.274 0.0042
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant E236A Bacillus subtilis
2.3.1.274 0.0043
-
 phosphate pH 7.5, 37°C, mutant E181A Bacillus subtilis
2.3.1.274 0.0045
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant E181A Bacillus subtilis
2.3.1.274 0.0045
-
 phosphate pH 7.5, 37°C, mutant E236A Bacillus subtilis
2.3.1.274 0.0067
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant K184R Bacillus subtilis
2.3.1.274 0.0068
-
 phosphate pH 7.5, 37°C, mutant K184R Bacillus subtilis
2.3.1.274 0.0103
-
 phosphate pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis
2.3.1.274 0.012
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant R114A Bacillus subtilis
2.3.1.274 0.012
-
 phosphate pH 7.5, 37°C, mutant R73A Bacillus subtilis
2.3.1.274 0.0125
-
 phosphate pH 7.5, 37°C, mutant N229Q Bacillus subtilis
2.3.1.274 0.0142
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant R73A Bacillus subtilis
2.3.1.274 0.0145
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant N229Q Bacillus subtilis
2.3.1.274 0.122
-
 phosphate pH 7.5, 37°C, mutant R114A Bacillus subtilis
2.3.1.274 0.123
-
 phosphate pH 7.5, 37°C, mutant R120A Bacillus subtilis
2.3.1.274 0.128
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, mutant R120A Bacillus subtilis
2.3.1.274 2.467
-
 holo-[acyl-carrier protein] pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis
2.3.1.274 2.783
-
 palmitoyl phosphate pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.274 7.5
-
 assay at Bacillus subtilis

General Information

EC Number General Information Comment Organism
2.3.1.274 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 [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.3.1.274 0.099
-
 phosphate pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis
2.3.1.274 0.907
-
 palmitoyl-[acyl-carrier protein] pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis
2.3.1.274 752.3
-
 palmitoyl phosphate pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis
2.3.1.274 6325.6
-
 holo-[acyl-carrier protein] pH 7.5, 37°C, recombinant wild-type enzyme Bacillus subtilis