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Literature summary for 3.1.1.1 extracted from

  • Manco, G.; Mandrich, L.; Rossi, M.
    Residues at the active site of the esterase 2 from Alicyclobacillus acidocaldarius involved in substrate specificity and catalytic activity at high temperature (2001), J. Biol. Chem., 276, 37482-37490.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of wild-type and mutants in Escherichia coli BL21(DE3) Alicyclobacillus acidocaldarius

Protein Variants

Protein Variants Comment Organism
M112S site-directed and saturation mutagenesis, altered substrate specificity, higher efficiency with monoacylesters whose chain length is greater than 8 C-atoms Alicyclobacillus acidocaldarius
M112S/R215L site-directed and saturation mutagenesis, altered substrate specificity, higher efficiency with monoacylesters whose chain length is greater than 8 C-atoms Alicyclobacillus acidocaldarius
M112T site-directed and saturation mutagenesis, altered substrate specificity Alicyclobacillus acidocaldarius
M112V site-directed and saturation mutagenesis, altered substrate specificity Alicyclobacillus acidocaldarius
R215L site-directed and saturation mutagenesis, altered substrate specificity, higher efficiency with monoacylesters whose chain length is greater than 8 C-atoms Alicyclobacillus acidocaldarius

Inhibitors

Inhibitors Comment Organism Structure
HEPES inhibition mechanism, competitive Alicyclobacillus acidocaldarius
propan-2-ol competitive Alicyclobacillus acidocaldarius

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.007
-
4-nitrophenyl dodecanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.008
-
4-nitrophenyl dodecanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.008
-
4-nitrophenyl decanoate mutants M211S and M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.01
-
4-nitrophenyl dodecanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.011
-
4-nitrophenyl decanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.015
-
4-nitrophenyl dodecanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.017
-
4-nitrophenyl tetradecanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.018
-
4-nitrophenyl decanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.019
-
4-nitrophenyl decanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.019
-
4-nitrophenyl tetradecanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.019
-
4-nitrophenyl dodecanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.02
-
4-nitrophenyl tetradecanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.021
-
4-nitrophenyl hexadecanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.021
-
4-nitrophenyl decanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.021
-
4-nitrophenyl dodecanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.022
-
4-nitrophenyl octanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.022
-
4-nitrophenyl tetradecanoate mutants R215L and M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.023
-
4-nitrophenyl hexadecanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.025
-
4-nitrophenyl octanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.026
-
4-nitrophenyl hexadecanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.028
-
4-nitrophenyl octanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.031
-
4-nitrophenyl hexadecanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.031
-
4-nitrophenyl tetradecanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.034
-
4-nitrophenyl hexadecanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.037
-
4-nitrophenyl octanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.042
-
4-nitrophenyl hexadecanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.06
-
4-nitrophenyl octanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.09
-
4-nitrophenyl hexanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.099
-
4-nitrophenyl hexanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.1
-
4-nitrophenyl octanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.21
-
4-nitrophenyl hexanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.22
-
4-nitrophenyl hexanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.41
-
4-nitrophenyl hexanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.51
-
4-nitrophenyl hexanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.73
-
4-nitrophenyl butanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
0.75
-
4-nitrophenyl butanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1.06
-
4-nitrophenyl butanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1.27
-
4-nitrophenyl butanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1.77
-
4-nitrophenyl butanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1.86
-
4-nitrophenyl butanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius

Organism

Organism UniProt Comment Textmining
Alicyclobacillus acidocaldarius
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutants from Escherichia coli BL21(DE3) Alicyclobacillus acidocaldarius

Reaction

Reaction Comment Organism Reaction ID
a carboxylic ester + H2O = an alcohol + a carboxylate reaction mechanism Alicyclobacillus acidocaldarius

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl acetate + H2O
-
Alicyclobacillus acidocaldarius 4-nitrophenol + acetate
-
r
4-nitrophenyl butanoate + H2O best substrate of mutants M211V, M211T, M211S and M211S/R215L Alicyclobacillus acidocaldarius 4-nitrophenol + butanoate
-
r
4-nitrophenyl decanoate + H2O
-
Alicyclobacillus acidocaldarius 4-nitrophenol + decanoate
-
r
4-nitrophenyl dodecanoate + H2O
-
Alicyclobacillus acidocaldarius 4-nitrophenol + dodecanoate
-
r
4-nitrophenyl hexadecanoate + H2O
-
Alicyclobacillus acidocaldarius 4-nitrophenol + hexadecanoate
-
r
4-nitrophenyl hexanoate + H2O best substrate of the wild-type enzyme Alicyclobacillus acidocaldarius 4-nitrophenol + hexanoate
-
r
4-nitrophenyl octanoate + H2O
-
Alicyclobacillus acidocaldarius 4-nitrophenol + octanoate
-
r
4-nitrophenyl tetradecanoate + H2O
-
Alicyclobacillus acidocaldarius 4-nitrophenol + tetradecanoate
-
r
additional information substrate specificities of wild-type and mutant enzymes, reverse reaction in performed in organic solvents Alicyclobacillus acidocaldarius ?
-
?

Synonyms

Synonyms Comment Organism
EST2
-
Alicyclobacillus acidocaldarius

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
-
Alicyclobacillus acidocaldarius

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
7
-
4-nitrophenyl hexadecanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
8.3
-
4-nitrophenyl hexadecanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
15.6
-
4-nitrophenyl hexadecanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
21
-
4-nitrophenyl hexadecanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
27
-
4-nitrophenyl tetradecanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
35
-
4-nitrophenyl hexadecanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
40
-
4-nitrophenyl tetradecanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
44
-
4-nitrophenyl hexadecanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
105
-
4-nitrophenyl tetradecanoate wild-type enzyme and mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
110
-
4-nitrophenyl dodecanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
112
-
4-nitrophenyl dodecanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
232
-
4-nitrophenyl tetradecanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
235
-
4-nitrophenyl tetradecanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
350
-
4-nitrophenyl dodecanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
390
-
4-nitrophenyl dodecanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
465
-
4-nitrophenyl decanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
626
-
4-nitrophenyl decanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
705
-
4-nitrophenyl dodecanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
860
-
4-nitrophenyl decanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
940
-
4-nitrophenyl dodecanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
950
-
4-nitrophenyl decanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
970
-
4-nitrophenyl decanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1376
-
4-nitrophenyl octanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1380
-
4-nitrophenyl octanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1500
-
4-nitrophenyl hexanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1645
-
4-nitrophenyl octanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1650
-
4-nitrophenyl octanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1670
-
4-nitrophenyl decanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1740
-
4-nitrophenyl octanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1855
-
4-nitrophenyl octanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1860
-
4-nitrophenyl octanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
1960
-
4-nitrophenyl octanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
2180
-
4-nitrophenyl butanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
2300
-
4-nitrophenyl hexanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
2750
-
4-nitrophenyl butanoate mutant M211S/R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
3080
-
4-nitrophenyl octanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
3200
-
4-nitrophenyl butanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
3420
-
4-nitrophenyl hexanoate wild-type enzyme, pH 7.1, 70°C Alicyclobacillus acidocaldarius
3660
-
4-nitrophenyl hexanoate mutant R215L, pH 7.1, 70°C Alicyclobacillus acidocaldarius
3890
-
4-nitrophenyl hexanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
4000
-
4-nitrophenyl butanoate mutant M211V, pH 7.1, 70°C Alicyclobacillus acidocaldarius
4230
-
4-nitrophenyl butanoate mutant M211S, pH 7.1, 70°C Alicyclobacillus acidocaldarius
4440
-
4-nitrophenyl hexanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius
5240
-
4-nitrophenyl butanoate mutant M211T, pH 7.1, 70°C Alicyclobacillus acidocaldarius

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.1
-
-
Alicyclobacillus acidocaldarius

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
130
-
propan-2-ol competitive, pH 7.1, 70°C Alicyclobacillus acidocaldarius
780
-
HEPES competitive, pH 7.1, 70°C Alicyclobacillus acidocaldarius