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Literature summary for 3.4.22.70 extracted from

  • Zhulenkovs, D.; Jaudzems, K.; Zajakina, A.; Leonchiks, A.
    Enzymatic activity of circular sortase A under denaturing conditions: an advanced tool for protein ligation (2014), Biochem. Eng. J., 82, 200-209.
No PubMed abstract available
Search for more literature for 3.4.22.70

Application

Application Comment Organism
additional information Staphylococcus aureus sortase A is a transpeptidase that is extensively used in various protein research applications, the enzyme is highly selective and does not require any cofactors for the catalysis of protein ligation and can be produced in high yields Staphylococcus aureus

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of linear truncated enzyme mutant LsrtADELTA59 in Escherichia coli strain BL21(DE3), development of an expression system in Escherichia coli that allows production of circularly closed sortase A truncated mutant CsrtADELTA59, overview. Efficient enzyme cyclization is achieved by Synechocystis sp. PCC6803 intein-mediated post-translational splicing, NMR spectroscopy of 15N-labeled LsrtADELTAN59 and CsrtADELTAN59 Staphylococcus aureus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.013
-
 Dabcyl-QALPETGEE-Edans pH 7.5, 37°C, recombinant linear truncated mutant LSrtADELTAN59 enzyme Staphylococcus aureus
0.014
-
 Dabcyl-QALPETGEE-Edans pH 7.5, 37°C, recombinant circular truncated mutant CSrtADELTAN59 enzyme Staphylococcus aureus

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ required Staphylococcus aureus

Organism

Organism UniProt Comment Textmining
Staphylococcus aureus
-
-
-
Staphylococcus aureus MSSA476
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant mutant and modified enzymes from Escherichia coli by cation and anion exchange chromatography Staphylococcus aureus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Dabcyl-QALPETGEE-Edans + H2O
-
 Staphylococcus aureus Dabcyl-QALPET + GEE-Edans
-
 ?
Dabcyl-QALPETGEE-Edans + H2O
-
 Staphylococcus aureus MSSA476 Dabcyl-QALPET + GEE-Edans
-
 ?
GST-LPETG + G-eGFP G-eGFP = enhanced green fluorescent protein with a glycine at the N-terminus, protein-protein ligation reaction Staphylococcus aureus ?
-
 ?
GST-LPETG + G-eGFP G-eGFP = enhanced green fluorescent protein with a glycine at the N-terminus, protein-protein ligation reaction Staphylococcus aureus MSSA476 ?
-
 ?
additional information the enzyme is highly selective and does not require any cofactors for the catalysis of protein ligation, but it is unable to access the recognition site within the highly structured regions of folded substrates. Ligation of purified glutathione-S-transferase and green fluorescence protein by recombinant circular enzyme Staphylococcus aureus ?
-
 ?
additional information the enzyme is highly selective and does not require any cofactors for the catalysis of protein ligation, but it is unable to access the recognition site within the highly structured regions of folded substrates. Ligation of purified glutathione-S-transferase and green fluorescence protein by recombinant circular enzyme Staphylococcus aureus MSSA476 ?
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Staphylococcus aureus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
125
-
 Dabcyl-QALPETGEE-Edans pH 7.5, 37°C, recombinant linear truncated mutant LSrtADELTAN59 enzyme Staphylococcus aureus
135
-
 Dabcyl-QALPETGEE-Edans pH 7.5, 37°C, recombinant circular truncated mutant CSrtADELTAN59 enzyme Staphylococcus aureus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Staphylococcus aureus

General Information

General Information Comment Organism
additional information the structural integrity of circular sortase A and its biochemical characteristics are compared to those of the linear enzyme analog and are similar under native conditions, overview. Circular enzyme is active at concentrations of urea up to 3 M and is capable of efficient catalytic protein-protein coupling, while the linear enzyme is unable to mediate the ligation of substrate proteins under the same conditions Staphylococcus aureus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
9615
-
 Dabcyl-QALPETGEE-Edans pH 7.5, 37°C, recombinant linear truncated mutant LSrtADELTAN59 enzyme Staphylococcus aureus
9642
-
 Dabcyl-QALPETGEE-Edans pH 7.5, 37°C, recombinant circular truncated mutant CSrtADELTAN59 enzyme Staphylococcus aureus