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Literature summary 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)

EC Number Cloned (Comment) Organism
3.1.1.1 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

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.1.1 69000
-
 or higher oligomeric state, 4 * 69000, calculation from amino acid sequence Pseudomonas sp.
3.1.1.1 300000
-
 recombinant enzyme, gel filtration Pseudomonas sp.

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.1 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)

EC Number Renatured (Comment) Organism
3.1.1.1 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]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.1.1 additional information
-
 substrate specificty Pseudomonas sp.
3.1.1.1 48
-
 purified renatured, recombinant enzyme, substrate p-tolyl octanoate Pseudomonas sp.
3.1.1.1 58.2
-
 purified renatured, recombinant enzyme, substrate 4-nitrophenyl butyrate Pseudomonas sp.
3.1.1.1 62
-
 purified renatured, recombinant enzyme, substrate vinyl octanoate Pseudomonas sp.
3.1.1.1 86
-
 purified renatured, recombinant enzyme, substrate vinyl hexanoate Pseudomonas sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.1 4-nitrophenyl acetate + H2O
-
 Pseudomonas sp. 4-nitrophenol + acetate
-
 ?
3.1.1.1 4-nitrophenyl acetate + H2O
-
 Pseudomonas sp. B11-1 4-nitrophenol + acetate
-
 ?
3.1.1.1 4-nitrophenyl butyrate + H2O
-
 Pseudomonas sp. 4-nitrophenol + butyrate
-
 ?
3.1.1.1 4-nitrophenyl butyrate + H2O
-
 Pseudomonas sp. B11-1 4-nitrophenol + butyrate
-
 ?
3.1.1.1 4-nitrophenyl caproate + H2O
-
 Pseudomonas sp. 4-nitrophenol + caproate
-
 ?
3.1.1.1 4-nitrophenyl caprylate + H2O
-
 Pseudomonas sp. 4-nitrophenol + caprylate
-
 ?
3.1.1.1 4-nitrophenyl laurate + H2O
-
 Pseudomonas sp. 4-nitrophenol + laurate
-
 ?
3.1.1.1 4-nitrophenyl palmitate + H2O
-
 Pseudomonas sp. 4-nitrophenol + palmitate
-
 ?
3.1.1.1 4-nitrophenyl valerate + H2O
-
 Pseudomonas sp. 4-nitrophenol + valerate
-
 ?
3.1.1.1 butyl octanoate + H2O
-
 Pseudomonas sp. butanol + octanoate
-
 ?
3.1.1.1 ethyl hexanoate + H2O
-
 Pseudomonas sp. ethanol + hexanoate
-
 ?
3.1.1.1 ethyl laurate + H2O
-
 Pseudomonas sp. ethanol + laurate
-
 ?
3.1.1.1 ethyl myristate + H2O
-
 Pseudomonas sp. ethanol + myristate
-
 ?
3.1.1.1 ethyl octanoate + H2O
-
 Pseudomonas sp. ethanol + octanoate
-
 ?
3.1.1.1 ethyl pelargonate + H2O
-
 Pseudomonas sp. ethanol + pelargonate
-
 ?
3.1.1.1 isopentyl octanoate + H2O
-
 Pseudomonas sp. isopentanol + octanoate
-
 ?
3.1.1.1 isopropyl octanoate + H2O
-
 Pseudomonas sp. isopropanol + octanoate
-
 ?
3.1.1.1 methyl octanoate + H2O
-
 Pseudomonas sp. methanol + octanoate
-
 ?
3.1.1.1 additional information hydrolysis of vinyl and aryl esters of chain length C4-C6 Pseudomonas sp. ?
-
 ?
3.1.1.1 additional information hydrolysis of vinyl and aryl esters of chain length C4-C6 Pseudomonas sp. B11-1 ?
-
 ?
3.1.1.1 p-tolyl octanoate + H2O
-
 Pseudomonas sp. p-toluol + octanoate
-
 ?
3.1.1.1 pentyl octanoate + H2O
-
 Pseudomonas sp. pentanol + octanoate
-
 ?
3.1.1.1 phenyl acetate + H2O
-
 Pseudomonas sp. phenol + acetate
-
 ?
3.1.1.1 phenyl acetate + H2O
-
 Pseudomonas sp. B11-1 phenol + acetate
-
 ?
3.1.1.1 tributyrin + H2O
-
 Pseudomonas sp. ?
-
 ?
3.1.1.1 tributyrin + H2O
-
 Pseudomonas sp. B11-1 ?
-
 ?
3.1.1.1 vinyl acetate + H2O
-
 Pseudomonas sp. vinyl alcohol + acetate
-
 ?
3.1.1.1 vinyl butyrate + H2O
-
 Pseudomonas sp. vinyl alcohol + butyrate
-
 ?
3.1.1.1 vinyl hexanoate + H2O
-
 Pseudomonas sp. vinyl alcohol + hexanoate
-
 ?
3.1.1.1 vinyl octanoate + H2O
-
 Pseudomonas sp. vinyl alcohol + octanoate
-
 ?
3.1.1.1 vinyl propionate + H2O
-
 Pseudomonas sp. vinyl alcohol + propionate
-
 ?

Subunits

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.1.1 additional information
-
 cold-adapted enzyme Pseudomonas sp.
3.1.1.1 45
-
 about Pseudomonas sp.

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.1.1.1 20 55 50% activity at 20°C and 55°C Pseudomonas sp.

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.1.1.1 30
-
 60 min, more than 90% remaining activity Pseudomonas sp.
3.1.1.1 50
-
 30 min, complete loss of activity Pseudomonas sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.1 8
-
 about Pseudomonas sp.