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Literature summary for 3.5.5.5 extracted from

  • Banerjee, A.; Dubey, S.; Kaul, P.; Barse, B.; Piotrowski, M.; Banerjee, U.C.
    enantioselective nitrilase from Pseudomonas putida: cloning, heterologous expression, and bioreactor studies (2008), Mol. Biotechnol., 41, 35-41.
    View publication on PubMed
Search for more literature for 3.5.5.5

Application

Application Comment Organism
biotechnology preparation of optically pure carboxylic acids Pseudomonas putida

Cloned(Commentary)

Cloned (Comment) Organism
The nitrilase gene from Pseudomonas putida is cloned in plasmid pET 21b(+) and over-expressed as histidine-tagged protein in Escherichia coli strain BL21 (DE3) Pseudomonas putida

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
45000
-
 determined by polyacrylamide gelelectrophorese Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
mandelonitrile + 2 H2O Pseudomonas putida racemic, enzyme hydrolizes the cyanide functionality to the corresponding carboxylic acid under ambient conditions (R)-mandelic acid + NH3
-
 ?
mandelonitrile + 2 H2O Pseudomonas putida MTCC 5110 racemic, enzyme hydrolizes the cyanide functionality to the corresponding carboxylic acid under ambient conditions (R)-mandelic acid + NH3
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas putida B0LAW4
-
-
Pseudomonas putida MTCC 5110 B0LAW4
-
-

Purification (Commentary)

Purification (Comment) Organism
cells of bacteria culture are disrupted and centrifugation and the enzyme in the soluble fraction is purified using Ni-NTA affinity chromatography Pseudomonas putida

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.44
-
 pH 8.0, in supernatant of the recombinant Escherichia coli cells, activity is highest after 6 hours of cultivation Pseudomonas putida
1.26
-
 pH 7.5, activity in enzyme preparation after purification, activity of the purified His-tagged protein was 28fold higher than the activity of the wild-type nitrilase Pseudomonas putida
15.14
-
 initial pH value 8.5 Pseudomonas putida
15.39
-
 initial pH value 6.5 Pseudomonas putida
16.03
-
 controlled pH value 6.5 Pseudomonas putida
16.27
-
 controlled pH value 8.5 Pseudomonas putida
18.05
-
 controlled pH value 7.5 Pseudomonas putida
18.92
-
 initial pH value 7.5 Pseudomonas putida
21.7
-
 Effect of IPG as inducer on the growth and enzyme production by Escherichia coli in a stirred tank reactor Pseudomonas putida
22.8
-
 Effect of lactose as inducer on the growth and enzyme production by Escherichia coli in a stirred tank reactor Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
mandelonitrile + 2 H2O racemic, enzyme hydrolizes the cyanide functionality to the corresponding carboxylic acid under ambient conditions Pseudomonas putida (R)-mandelic acid + NH3
-
 ?
mandelonitrile + 2 H2O racemic, enzyme hydrolizes the cyanide functionality to the corresponding carboxylic acid under ambient conditions Pseudomonas putida MTCC 5110 (R)-mandelic acid + NH3
-
 ?

Synonyms

Synonyms Comment Organism
enantioselective arylacetonitrilase
-
 Pseudomonas putida
nitrilase
-
 Pseudomonas putida

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 in fermentation Pseudomonas putida