Activating Compound | Comment | Organism | Structure |
---|---|---|---|
NHase activator protein | the activator gene encding activator protein might be involved in protein folding of the alpha- and beta-subunits of NHase | Pseudomonas putida |
Cloned (Comment) | Organism |
---|---|
gene Pput_2728, gene cluster and phylogenetic analysis, functional expression of His6-tagged subunits in Escherichia coli strain BL21(DE3) by assisting of a putative activator gene adjacent to beta-subunit region. Maximal enzyme activity is obtained when the structural and activator genes are transcribed as one unit in plasmid pCDFDuet-1 at 18°C. The expressed product does not show any NHase activity when the downstream activator gene is ignored, and the product completely exists in insoluble inclusion body | Pseudomonas putida |
gene Pput_2729, gene cluster and phylogenetic analysis, functional expression of His6-tagged subunits in Escherichia coli strain BL21(DE3) by assisting of a putative activator gene adjacent to beta-subunit region. Maximal enzyme activity is obtained when the structural and activator genes are transcribed as one unit in plasmid pCDFDuet-1 at 18°C. The expressed product does not show any NHase activity when the downstream activator gene is ignored, and the product completely exists in insoluble inclusion body | Pseudomonas putida |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | Fe-type NHase | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | A5W402 | gene Pput_2728 encoding subunit alpha | - |
Pseudomonas putida | A5W402 | gene Pput_2729 encoding subunit alpha | - |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged subunits 2.6fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and desalting gel filtration | Pseudomonas putida |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
26.61 | - |
purified recombinant enzyme, pH 8.0, 25°C, substrate 3-cyanopyridine | Pseudomonas putida |
535 | - |
purified recombinant enzyme, pH 8.0, 25°C, substrate valeronitrile | Pseudomonas putida |
553 | - |
purified recombinant enzyme, pH 8.0, 25°C, substrate 4-chlorobutyronitrile | Pseudomonas putida |
775 | - |
purified recombinant enzyme, pH 8.0, 25°C, substrate isobutyronitrile | Pseudomonas putida |
941 | - |
purified recombinant enzyme, pH 8.0, 25°C, substrate acrylonitrile | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-cyanopyridine + H2O | lower activity | Pseudomonas putida | pyridine-3-carbamide | - |
? | |
3-methylbenzonitrile + H2O | lower activity | Pseudomonas putida | 3-methylbenzamide | - |
? | |
4-chlorobutyronitrile + H2O | - |
Pseudomonas putida | 4-chlorobutyramide | - |
? | |
4-cyanopyridine + H2O | lower activity | Pseudomonas putida | isonicotinamide | - |
? | |
acrylonitrile + H2O | best substrate | Pseudomonas putida | acrylamide | - |
? | |
benzeneacetonitrile + H2O | very low activity | Pseudomonas putida | ? | - |
? | |
benzonitrile + H2O | lower activity | Pseudomonas putida | benzamide | - |
? | |
isobutyronitrile + H2O | - |
Pseudomonas putida | isobutyramide | - |
? | |
additional information | the Fe-type NHase exhibits broad substrate specificity | Pseudomonas putida | ? | - |
? | |
valeronitrile + H2O | - |
Pseudomonas putida | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NHase | - |
Pseudomonas putida |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | - |
- |
Pseudomonas putida |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
10 | 45 | activity range, profile overview | Pseudomonas putida |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | - |
purified recombinant enzyme, over 60% activity remaining after 3 h | Pseudomonas putida |
20 | 45 | purified recombinant enzyme, 50% of maximal activity within this range. Half-lives of NHase F1 at 20°C, 25°C, 30°C, and 35°C are calculated as 5.1 h, 3.8 h, 2.2 h, and 1.2 h, respectively | Pseudomonas putida |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Pseudomonas putida |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 10 | activity range, profile overview | Pseudomonas putida |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | 8 | purified recombinant enzyme, 25°C, highest stability at | Pseudomonas putida |
General Information | Comment | Organism |
---|---|---|
evolution | because of the absence of sequence similarity between the Co- and Fe-type NHase activators, the two types of NHases might be assembled and maturated by different molecular mechanisms | Pseudomonas putida |