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Literature summary for 2.3.1.28 extracted from

  • Seo, H.; Lee, J.W.; Garcia, S.; Trinh, C.T.
    Single mutation at a highly conserved region of chloramphenicol acetyltransferase enables isobutyl acetate production directly from cellulose by Clostridium thermocellum at elevated temperatures (2019), Biotechnol. Biofuels, 12, 245 .
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
synthesis engineering of a thermostable chloramphenicol acetyltransferase for enhanced isobutyl acetate production at elevated temperatures. Mutant F97W significantly increases conversion of isobutanol to isobutyl acetate. Using F97W, direct conversion of cellulose into isobutyl acetate is possible by an engineered Clostridium thermocellum strain at elevated temperatures Staphylococcus aureus

Crystallization (Commentary)

Crystallization (Comment) Organism
homology modeling of structure. The binding pocket consists of Tyr20, Phe27, Tyr50, Thr88, Ile89, Phe90, Phe97, Ser140, Leu141, Ser142, Ile143, Ile144, Pro145, Trp146, Phe152, Leu154, Ile166, Ile167, Thr168, His189, Asp193, Gly194, and Tyr195, where His189 and Asp193 are the catalytic sites Staphylococcus aureus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
chloramphenicol mutant F97W, pH 8.0, 50°C Staphylococcus aureus
0.28
-
chloramphenicol wild-type, pH 8.0, 50°C Staphylococcus aureus
138.7
-
isobutanol wild-type, pH 8.0, 50°C Staphylococcus aureus
144.8
-
isobutanol mutant F97W, pH 8.0, 50°C Staphylococcus aureus

Organism

Organism UniProt Comment Textmining
Staphylococcus aureus
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetyl-CoA + 2-phenylethanol
-
Staphylococcus aureus CoA + 2-phenylethyl acetate
-
?
acetyl-CoA + benzyl alcohol
-
Staphylococcus aureus CoA + benzyl acetate
-
?
acetyl-CoA + chloramphenicol
-
Staphylococcus aureus CoA + chloramphenicol 3-acetate
-
?
acetyl-CoA + isobutanol
-
Staphylococcus aureus CoA + isobutyl acetate
-
?
additional information poor substrates: butanol, propanol, ethanol, isopropanol. Specific ester production titers and rates are higher for aromatic alcohols than linear, short-chain alcohols Staphylococcus aureus ?
-
-

Subunits

Subunits Comment Organism
? x * 25800, SDS-PAGE Staphylococcus aureus

Synonyms

Synonyms Comment Organism
CAT
-
Staphylococcus aureus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
68.3
-
melting temperature, mutant F97W Staphylococcus aureus
72
-
melting temperature, wild-type Staphylococcus aureus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.3
-
isobutanol wild-type, pH 8.0, 50°C Staphylococcus aureus
0.6
-
isobutanol mutant F97W, pH 8.0, 50°C Staphylococcus aureus
102.6
-
chloramphenicol mutant F97W, pH 8.0, 50°C Staphylococcus aureus
203
-
chloramphenicol wild-type, pH 8.0, 50°C Staphylococcus aureus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0022
-
isobutanol wild-type, pH 8.0, 50°C Staphylococcus aureus
0.0041
-
isobutanol mutant F97W, pH 8.0, 50°C Staphylococcus aureus
577
-
chloramphenicol mutant F97W, pH 8.0, 50°C Staphylococcus aureus
737
-
chloramphenicol wild-type, pH 8.0, 50°C Staphylococcus aureus