EC Number | Cloned (Comment) | Organism |
---|---|---|
1.13.11.66 | genes pnpCD, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Pseudomonas sp. |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.13.11.66 | purified full-length or proteolytically truncated PnpCD in apo form and in complex with Fe3+ or substrate analogue hydroxybenzonitrile and Cd2+, i.e. apo-PnpCD, PnpCD-Fe3+, and PnpCD-Cd2+-HBN, sitting-drop vapor diffusion method, 15-20 mg/ml protein in 10mM Tris-HCl, pH 8.0, and 100 mM NaCl, is mixed with reservoir solution containing 0.2 M sodium thiocyanate, 20% w/v PEG 3350, 20°C, method optimization, X-ray diffraction structure determination and analysis | Pseudomonas sp. |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.13.11.66 | E248Q | site-directed mutagenesis, the mutation results in complete loss of enzyme activity | Pseudomonas sp. |
1.13.11.66 | F264A | site-directed mutagenesis, the mutation results in almost complete loss of enzyme activity | Pseudomonas sp. |
1.13.11.66 | F79A | site-directed mutagenesis, the mutation results in almost complete loss of enzyme activity | Pseudomonas sp. |
1.13.11.66 | L252A | site-directed mutagenesis, the mutation results in a 70% loss of enzyme activity | Pseudomonas sp. |
1.13.11.66 | L313A | site-directed mutagenesis, the mutation results in almost complete loss of enzyme activity | Pseudomonas sp. |
1.13.11.66 | V315A | site-directed mutagenesis, the mutation results in a 50% loss of enzyme activity | Pseudomonas sp. |
1.13.11.66 | W230A | site-directed mutagenesis, the mutation results in a 70% loss of enzyme activity | Pseudomonas sp. |
1.13.11.66 | W273a | site-directed mutagenesis, the mutation results in almost complete loss of enzyme activity | Pseudomonas sp. |
1.13.11.66 | W76A | site-directed mutagenesis, the mutation results in almost complete loss of enzyme activity | Pseudomonas sp. |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.13.11.66 | Fe2+ | a Fe2+-dependent dioxygenase, selectively utilizes Fe2+for its catalytic reaction. Four residues of enzyme PnpD, His256, Asn258, Glu262, and His303, coordinate the iron ion | Pseudomonas sp. | |
1.13.11.66 | Fe3+ | binding structure, overview | Pseudomonas sp. | |
1.13.11.66 | additional information | Fe3+, Mn2+, Co2+, Ni2+, Cu2+, and Cd2+ cannot substitute for Fe2+ | Pseudomonas sp. |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.13.11.66 | 4-nitrophenol + O2 | Pseudomonas sp. | - |
? | - |
? | |
1.13.11.66 | benzene-1,4-diol + O2 | Pseudomonas sp. | - |
(2Z,4E)-4-hydroxy-6-oxohexa-2,4-dienoate | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.13.11.66 | Pseudomonas sp. | C1I210 AND C1I209 | beta- and alpha-subunit | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.13.11.66 | recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Pseudomonas sp. |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.13.11.66 | 4-nitrophenol + O2 | - |
Pseudomonas sp. | ? | - |
? | |
1.13.11.66 | benzene-1,4-diol + O2 | - |
Pseudomonas sp. | (2Z,4E)-4-hydroxy-6-oxohexa-2,4-dienoate | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.13.11.66 | heterotetramer | 2 * 38300, subunit alpha, + 2 * 18000, subunit beta | Pseudomonas sp. |
1.13.11.66 | More | the PnpCD structure contains a pseudo cupin and an iron metallocenter in the catalytic PnpD. Both the PnpC and the C-terminal domains of PnpD comprise a conserved cupin fold, whereas PnpC cannot form a competent metal binding pocket as can PnpD cupin, structure analysis, overview | Pseudomonas sp. |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.13.11.66 | PnpCD | - |
Pseudomonas sp. |
1.13.11.66 | two-subunit hydroquinone 1,2-dioxygenase | - |
Pseudomonas sp. |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.13.11.66 | 25 | - |
assay at | Pseudomonas sp. |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.13.11.66 | 7 | - |
assay at | Pseudomonas sp. |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.13.11.66 | metabolism | the two-subunit hydroquinone 1,2-dioxygenase PnpCD is the ring cleavage enzyme in para-nitrophenol catabolism | Pseudomonas sp. |
1.13.11.66 | additional information | the PnpCD structure contains a pseudo cupin and a iron metallocenter in the catalytic PnpD, which adds to understanding of the ring cleavage mechanism of dioxygenases, structure analysis, overview | Pseudomonas sp. |
1.13.11.66 | physiological function | hydroquinone 1,2-dioxygenase PnpCD is the key enzyme in the hydroquinone pathway of para-nitrophenol degradation, catalyzes the ring cleavage of hydroquinone to gamma-hydroxymuconic semialdehyde | Pseudomonas sp. |