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Literature summary for 3.1.1.1 extracted from

  • Suzuki, T.; Nakayama, T.; Choo, D.W.; Hirano, Y.; Kurihara, T.; Nishino, T.; Esaki, N.
    Cloning, heterologous expression, renaturation, and characterization of a cold-adapted esterase with unique primary structure from a psychrotroph Pseudomonas sp. strain B11-1 (2003), Protein Expr. Purif., 30, 171-178.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene PsEst1, DNA and amino acid sequence determination and analysis, overexpression of the cold-adapted enzyme in Escherichia coli BL21(DE3), formation of inclusion bodies Pseudomonas sp.

General Stability

General Stability Organism
Triton X-100 is essential for enzyme stability after solubilization Pseudomonas sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
69000
-
or higher oligomeric state, 4 * 69000, calculation from amino acid sequence Pseudomonas sp.
300000
-
recombinant enzyme, gel filtration Pseudomonas sp.

Organism

Organism UniProt Comment Textmining
Pseudomonas sp. Q8GR22 cold-adapted enzyme
-
Pseudomonas sp. B11-1 Q8GR22 cold-adapted enzyme
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme solubilized as denatured protein from inclusion bodies after expression in Escherichia coli, denaturation by 8 M urea, purification to homogeneity Pseudomonas sp.

Renatured (Commentary)

Renatured (Comment) Organism
renaturation to the catalytically active recombinant enzyme from 8 M urea by dialysis in presence of 0.1% Triton X-100, detergents are essential for the efficient renaturation, 0.05-0.1% Triton X-100 is most efficient followed by 0.1% cholic acid, 30% glycerol and 0.05% CHAPS, and 0.1 M sarcosine, overview Pseudomonas sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
substrate specificty Pseudomonas sp.
48
-
purified renatured, recombinant enzyme, substrate p-tolyl octanoate Pseudomonas sp.
58.2
-
purified renatured, recombinant enzyme, substrate 4-nitrophenyl butyrate Pseudomonas sp.
62
-
purified renatured, recombinant enzyme, substrate vinyl octanoate Pseudomonas sp.
86
-
purified renatured, recombinant enzyme, substrate vinyl hexanoate Pseudomonas sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl acetate + H2O
-
Pseudomonas sp. 4-nitrophenol + acetate
-
?
4-nitrophenyl acetate + H2O
-
Pseudomonas sp. B11-1 4-nitrophenol + acetate
-
?
4-nitrophenyl butyrate + H2O
-
Pseudomonas sp. 4-nitrophenol + butyrate
-
?
4-nitrophenyl butyrate + H2O
-
Pseudomonas sp. B11-1 4-nitrophenol + butyrate
-
?
4-nitrophenyl caproate + H2O
-
Pseudomonas sp. 4-nitrophenol + caproate
-
?
4-nitrophenyl caprylate + H2O
-
Pseudomonas sp. 4-nitrophenol + caprylate
-
?
4-nitrophenyl laurate + H2O
-
Pseudomonas sp. 4-nitrophenol + laurate
-
?
4-nitrophenyl palmitate + H2O
-
Pseudomonas sp. 4-nitrophenol + palmitate
-
?
4-nitrophenyl valerate + H2O
-
Pseudomonas sp. 4-nitrophenol + valerate
-
?
butyl octanoate + H2O
-
Pseudomonas sp. butanol + octanoate
-
?
ethyl hexanoate + H2O
-
Pseudomonas sp. ethanol + hexanoate
-
?
ethyl laurate + H2O
-
Pseudomonas sp. ethanol + laurate
-
?
ethyl myristate + H2O
-
Pseudomonas sp. ethanol + myristate
-
?
ethyl octanoate + H2O
-
Pseudomonas sp. ethanol + octanoate
-
?
ethyl pelargonate + H2O
-
Pseudomonas sp. ethanol + pelargonate
-
?
isopentyl octanoate + H2O
-
Pseudomonas sp. isopentanol + octanoate
-
?
isopropyl octanoate + H2O
-
Pseudomonas sp. isopropanol + octanoate
-
?
methyl octanoate + H2O
-
Pseudomonas sp. methanol + octanoate
-
?
additional information hydrolysis of vinyl and aryl esters of chain length C4-C6 Pseudomonas sp. ?
-
?
additional information hydrolysis of vinyl and aryl esters of chain length C4-C6 Pseudomonas sp. B11-1 ?
-
?
p-tolyl octanoate + H2O
-
Pseudomonas sp. p-toluol + octanoate
-
?
pentyl octanoate + H2O
-
Pseudomonas sp. pentanol + octanoate
-
?
phenyl acetate + H2O
-
Pseudomonas sp. phenol + acetate
-
?
phenyl acetate + H2O
-
Pseudomonas sp. B11-1 phenol + acetate
-
?
tributyrin + H2O
-
Pseudomonas sp. ?
-
?
tributyrin + H2O
-
Pseudomonas sp. B11-1 ?
-
?
vinyl acetate + H2O
-
Pseudomonas sp. vinyl alcohol + acetate
-
?
vinyl butyrate + H2O
-
Pseudomonas sp. vinyl alcohol + butyrate
-
?
vinyl hexanoate + H2O
-
Pseudomonas sp. vinyl alcohol + hexanoate
-
?
vinyl octanoate + H2O
-
Pseudomonas sp. vinyl alcohol + octanoate
-
?
vinyl propionate + H2O
-
Pseudomonas sp. vinyl alcohol + propionate
-
?

Subunits

Subunits Comment Organism
tetramer or higher oligomeric state, 4 * 69000, calculation from amino acid sequence Pseudomonas sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
additional information
-
cold-adapted enzyme Pseudomonas sp.
45
-
about Pseudomonas sp.

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 55 50% activity at 20°C and 55°C Pseudomonas sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30
-
60 min, more than 90% remaining activity Pseudomonas sp.
50
-
30 min, complete loss of activity Pseudomonas sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
about Pseudomonas sp.