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Literature summary for 3.1.1.1 extracted from

  • Jeon, J.H.; Kim, S.J.; Lee, H.S.; Cha, S.S.; Lee, J.H.; Yoon, S.H.; Koo, B.S.; Lee, C.M.; Choi, S.H.; Lee, S.H.; Kang, S.G.; Lee, J.H.
    A novel metagenome-derived carboxylesterase that hydrolyzes beta-lactam antibiotics (2011), Appl. Environ. Microbiol., 77, 7830-7836.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells uncultured bacterium

Inhibitors

Inhibitors Comment Organism Structure
cephaloridine competitive inhibitor of 4-nitrophenyl butyrate uncultured bacterium
cephalothin competitive inhibitor of 4-nitrophenyl butyrate uncultured bacterium
additional information EstU1 activity is not affected by the presence of EDTA (94% activity at 1 mM) uncultured bacterium
phenylmethylsulfonyl fluoride EstU1 loses approximately 70% of its activity in the presence of 1 mM phenylmethylsulfonyl fluoride uncultured bacterium

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00603
-
4-nitrophenyl butyrate at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium
0.07849
-
cefazolin at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium

Metals/Ions

Metals/Ions Comment Organism Structure
additional information EstU1 activity is not affected by the presence of Ca2+ (91% activity at 1 mM), Co2+ (89% activity at 1 mM), Cu2+ (88% activity at 1 mM), Fe2+ (84% activity at 1 mM), Mg2+ (91% activity at 1 mM), Mn2+ (74% activity at 1 mM), Ni2+ (89% activity at 1 mM), and Zn2+ (78% activity at 1 mM) uncultured bacterium

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
44000
-
x * 44000, SDS-PAGE uncultured bacterium

Organism

Organism UniProt Comment Textmining
uncultured bacterium
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Talon metal affinity resin column chromatography and Superdex-75 gel filtration uncultured bacterium

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.75
-
using 4-nitrophenyl decanoate as substrate, at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium
6.24
-
using 4-nitrophenyl acetate as substrate, at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium
7.46
-
using 4-nitrophenyl octanoate as substrate, at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium
11.78
-
using 4-nitrophenyl hexanoate as substrate, at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium
22.24
-
using 4-nitrophenyl butyrate as substrate, at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl acetate + H2O
-
uncultured bacterium 4-nitrophenol + acetate
-
?
4-nitrophenyl butyrate + H2O highest activity uncultured bacterium 4-nitrophenol + butyrate
-
?
4-nitrophenyl decanoate + H2O
-
uncultured bacterium 4-nitrophenol + decanoate
-
?
4-nitrophenyl hexanoate + H2O
-
uncultured bacterium 4-nitrophenol + hexanoate
-
?
4-nitrophenyl octanoate + H2O
-
uncultured bacterium 4-nitrophenol + octanoate
-
?
ampicillin + H2O
-
uncultured bacterium ?
-
?
cefazolin + H2O
-
uncultured bacterium (6R,7R)-7-amino-3-{[(5-methyl-1,3,4-thiadiazol-2-yl)sulfanyl]methyl}-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid + (1H-tetrazol-1-yl)acetic acid
-
?
cephaloridine + H2O
-
uncultured bacterium (6R,7R)-7-amino-8-oxo-3-[(pyridin-1-ium-1-yl)methyl]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate + (thiophen-2-yl)acetic acid
-
?
cephalothin + H2O
-
uncultured bacterium acetate + deacetylcephalothin
-
?
additional information no enzyme activity is detected toward 4-nitrophenyl laurate, 4-nitrophenyl myristate, 4-nitrophenyl palmitate, and 4-nitrophenyl stearate uncultured bacterium ?
-
?
nitrocefin + H2O i.e. 3-(2,4-dinitrosyl)-(6R,7R)-7-(2-thienylacetamido)-ceph-3-em-4-carboxylic acid uncultured bacterium ?
-
?
penicillin G + H2O
-
uncultured bacterium 6-aminopenicillanic acid + phenylacetic acid
-
?

Subunits

Subunits Comment Organism
? x * 44000, SDS-PAGE uncultured bacterium

Synonyms

Synonyms Comment Organism
EstU1
-
uncultured bacterium

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
-
uncultured bacterium

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
5 75
-
uncultured bacterium

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.000238
-
cefazolin at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium
15.72
-
4-nitrophenyl butyrate at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
-
uncultured bacterium

pH Range

pH Minimum pH Maximum Comment Organism
7.5 9.5
-
uncultured bacterium

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.14
-
cephalothin at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium
0.215
-
cephaloridine at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.003035
-
cefazolin at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium
2608
-
4-nitrophenyl butyrate at 35°C in 100 mM sodium phosphate (pH 7.0) uncultured bacterium