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Literature summary for 2.3.1.274 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
Search for more literature for 2.3.1.274

Cloned(Commentary)

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

Protein Variants

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Bacillus subtilis P71018
-
-
Bacillus subtilis 168 P71018
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

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

Turnover Number [1/s]

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

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

General Information

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 [mM/s]

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