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Literature summary for 6.1.1.7 extracted from

  • De Pascale, G.; Lloyd, A.J.; Schouten, J.A.; Gilbey, A.M.; Roper, D.I.; Dowson, C.G.; Bugg, T.D.
    Kinetic characterisation of lipid II-Ala:alanyl tRNA ligase (MurN) from Streptococcus pneumoniae using semi-synthetic aminoacyl-lipid II substrates (2008), J. Biol. Chem., 283, 34571-34579.
    View publication on PubMedView publication on EuropePMC
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Application

Application Comment Organism
molecular biology a combined action of ligases MurM and MurN is required in order to rationalise the high level of dipeptide cross-links in penicillin-resistant Streptococcus pneumoniae, with ligase MurM showing the major difference between penicillin-resistant and penicillin-sensitive strains Streptococcus pneumoniae

Cloned(Commentary)

Cloned (Comment) Organism
recombinant MurN is expressed and purified as MBP fusion protein in Escherichia coli Streptococcus pneumoniae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0331
-
 lipid II L-Ala strain 159 Streptococcus pneumoniae
0.04
-
 lipid II L-Ala strain Pn16 Streptococcus pneumoniae
0.125
-
 lipid II L-Ser strain Pn16 Streptococcus pneumoniae
0.146
-
 lipid II L-Ser strain 159 Streptococcus pneumoniae

Organism

Organism UniProt Comment Textmining
Streptococcus pneumoniae
-
 strains 159, Pn16
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-Sepharose columns and anion-exchange chromatography. The yield of purified protein is 4-5 mg of protein per litre of culture, judged to be above 95% purity by SDS-PAGE Streptococcus pneumoniae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0007
-
 substrate: lipid II L-Ser, strain 159 Streptococcus pneumoniae
0.0009
-
 substrate: lipid II L-Ser, strain Pn16 Streptococcus pneumoniae
0.0087
-
 substrate: lipid II L-Ala, strain Pn16 Streptococcus pneumoniae
0.011
-
 substrate: lipid II L-Ala, strain 159 Streptococcus pneumoniae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-alanine + tRNAAla
-
 Streptococcus pneumoniae AMP + diphosphate + L-alanyl-tRNAAla
-
 ?
ATP + lipid II L-alanine + tRNAAla
-
 Streptococcus pneumoniae ?
-
 ?
ATP + lipid II L-serine + tRNAAla
-
 Streptococcus pneumoniae ?
-
 ?

Synonyms

Synonyms Comment Organism
alanyl tRNA ligase
-
 Streptococcus pneumoniae
MurN
-
 Streptococcus pneumoniae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Streptococcus pneumoniae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0166
-
 lipid II L-Ser strain 159 Streptococcus pneumoniae
0.0166
-
 lipid II L-Ser strain Pn16 Streptococcus pneumoniae
0.0635
-
 lipid II L-Ala strain Pn16 Streptococcus pneumoniae
0.071
-
 lipid II L-Ala strain 159 Streptococcus pneumoniae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.8
-
 assay at Streptococcus pneumoniae