EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.1.214 | gene cpr-c2, recombinant expression of His-tagged enzyme in Escherichia coli strain Rosetta (DE3) | Candida parapsilosis |
1.1.1.358 | expressed in Escherichia coli Rosetta (DE3) cells | Candida parapsilosis |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.1.1.214 | purified enzyme in apoform and in complex with NADPH, mixing of 0.001 ml of 15 mg/ml protein in 20 mM Tris-HCl, pH 8.0, with 0.001 ml of reservoir solution consisting of 0.1 M Tris-HCl, pH 8.1, and 23% w/v PEG 3350 at 20°C, for the enzyme-NADPH complexed crystals, the protein in 20 mM Tris-HCl, pH 8.0, and 5 mM MADPH, is mixed with 0.1 M TrisHCl, pH 7.4, and 256% w/v PEG 3350, X-ray diffraction structure determination and analysis at 1.70 A and 1.80 A resolution, respectively, molecular replacement method | Candida parapsilosis |
1.1.1.358 | apoenzyme and in complex with NADPH, sitting drop vapor diffusion method, using 0.1 M TrisHCl (pH 8.1) and 23% (w/v) polyethylene glycol 3350, at 20°C | Candida parapsilosis |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.1.1.214 | D58A | site-directed mutagenesis, the mutant shows 4.82% of wild-type activity | Candida parapsilosis |
1.1.1.214 | F299A | site-directed mutagenesis, the mutant shows 19.1% of wild-type activity | Candida parapsilosis |
1.1.1.214 | F300A | site-directed mutagenesis, the mutant shows 17.8% of wild-type activity | Candida parapsilosis |
1.1.1.214 | H125A | site-directed mutagenesis, the mutant shows 1.5% of wild-type activity | Candida parapsilosis |
1.1.1.214 | K264A | site-directed mutagenesis, the mutant shows 65.7% of wild-type activity | Candida parapsilosis |
1.1.1.214 | K28A | site-directed mutagenesis, the mutant shows 71.1% of wild-type activity | Candida parapsilosis |
1.1.1.214 | K30A | site-directed mutagenesis, the mutant shows 55.1% of wild-type activity | Candida parapsilosis |
1.1.1.214 | K88A | site-directed mutagenesis, inactive mutant | Candida parapsilosis |
1.1.1.214 | R267A | site-directed mutagenesis, the mutant shows 8.43% of wild-type activity | Candida parapsilosis |
1.1.1.214 | T27A | site-directed mutagenesis, the mutant shows 5.75% of wild-type activity | Candida parapsilosis |
1.1.1.214 | Y63A | site-directed mutagenesis, the mutant shows 0.17% of wild-type activity | Candida parapsilosis |
1.1.1.358 | D58A | 4.82% activity compared to the wild type enzyme | Candida parapsilosis |
1.1.1.358 | F299A | 19.1% activity compared to the wild type enzyme | Candida parapsilosis |
1.1.1.358 | F300A | 17.8% activity compared to the wild type enzyme | Candida parapsilosis |
1.1.1.358 | H125A | 1.5% activity compared to the wild type enzyme | Candida parapsilosis |
1.1.1.358 | K264A | 65.7% activity compared to the wild type enzyme | Candida parapsilosis |
1.1.1.358 | K28A | 71.1% activity compared to the wild type enzyme | Candida parapsilosis |
1.1.1.358 | K30A | 55.1% activity compared to the wild type enzyme | Candida parapsilosis |
1.1.1.358 | K88A | inactive | Candida parapsilosis |
1.1.1.358 | R267A | 8.43% activity compared to the wild type enzyme | Candida parapsilosis |
1.1.1.358 | T27A | 5.75% activity compared to the wild type enzyme | Candida parapsilosis |
1.1.1.358 | Y63A | 0.17% activity compared to the wild type enzyme | Candida parapsilosis |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.214 | (R)-pantolactone + NADP+ | Candida parapsilosis | - |
2-dehydropantolactone + NADPH + H+ | - |
r | |
1.1.1.214 | (R)-pantolactone + NADP+ | Candida parapsilosis IFO 0708 | - |
2-dehydropantolactone + NADPH + H+ | - |
r | |
1.1.1.214 | 2-dehydropantolactone + NADPH + H+ | Candida parapsilosis | the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner | (R)-pantolactone + NADP+ | - |
r | |
1.1.1.214 | 2-dehydropantolactone + NADPH + H+ | Candida parapsilosis IFO 0708 | the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner | (R)-pantolactone + NADP+ | - |
r | |
1.1.1.358 | 2-dehydropantolactone + NADPH + H+ | Candida parapsilosis | - |
(R)-pantolactone + NADP+ | - |
ir | |
1.1.1.358 | 2-dehydropantolactone + NADPH + H+ | Candida parapsilosis IFO 0708 | - |
(R)-pantolactone + NADP+ | - |
ir |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.1.214 | Candida parapsilosis | Q76L36 | - |
- |
1.1.1.214 | Candida parapsilosis IFO 0708 | Q76L36 | - |
- |
1.1.1.358 | Candida parapsilosis | Q76L36 | - |
- |
1.1.1.358 | Candida parapsilosis IFO 0708 | Q76L36 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.1.1.214 | recombinant His-tagged enzyme from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography, and tag cleavage by thrombin, followed by anion exchange chromatography, gel filtration, and dialysis | Candida parapsilosis |
1.1.1.358 | Ni Sepharose column chromatography, Resource Q column chromatography, and Superdex 75 gel filtration | Candida parapsilosis |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
1.1.1.214 | 23.6 | - |
purified recombinant detagged wild-type enzyme, pH and temperature not specified in the publication | Candida parapsilosis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.214 | (R)-pantolactone + NADP+ | - |
Candida parapsilosis | 2-dehydropantolactone + NADPH + H+ | - |
r | |
1.1.1.214 | (R)-pantolactone + NADP+ | - |
Candida parapsilosis IFO 0708 | 2-dehydropantolactone + NADPH + H+ | - |
r | |
1.1.1.214 | 2-dehydropantolactone + NADPH + H+ | the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner | Candida parapsilosis | (R)-pantolactone + NADP+ | - |
r | |
1.1.1.214 | 2-dehydropantolactone + NADPH + H+ | the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner | Candida parapsilosis IFO 0708 | (R)-pantolactone + NADP+ | - |
r | |
1.1.1.214 | additional information | structural basis of the substrate specificity, overview. Enzyme CPR-C2 adopts a triose-phosphate isomerase barrel fold at the core of the structure. Binding with the cofactor NADPH induces conformational changes in which Thr27 and Lys28 move 15 and 5.0 A, respectively, in the close vicinity of the adenosine 2'-phosphate group of NADPH to form hydrogen bonds, substrate binding modeling | Candida parapsilosis | ? | - |
? | |
1.1.1.214 | additional information | structural basis of the substrate specificity, overview. Enzyme CPR-C2 adopts a triose-phosphate isomerase barrel fold at the core of the structure. Binding with the cofactor NADPH induces conformational changes in which Thr27 and Lys28 move 15 and 5.0 A, respectively, in the close vicinity of the adenosine 2'-phosphate group of NADPH to form hydrogen bonds, substrate binding modeling | Candida parapsilosis IFO 0708 | ? | - |
? | |
1.1.1.358 | 2-dehydropantolactone + NADPH + H+ | - |
Candida parapsilosis | (R)-pantolactone + NADP+ | - |
ir | |
1.1.1.358 | 2-dehydropantolactone + NADPH + H+ | - |
Candida parapsilosis IFO 0708 | (R)-pantolactone + NADP+ | - |
ir |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.1.1.214 | conjugated polyketone reductase C2 | - |
Candida parapsilosis |
1.1.1.214 | CPR-C2 | - |
Candida parapsilosis |
1.1.1.214 | NADPH-dependent ketopantoyl lactone reductase | - |
Candida parapsilosis |
1.1.1.214 | nicotinamide adenine dinucleotide phosphate-dependent ketopantoyl lactone reductase | - |
Candida parapsilosis |
1.1.1.358 | conjugated polyketone reductase C2 | - |
Candida parapsilosis |
1.1.1.358 | CPR-C2 | - |
Candida parapsilosis |
1.1.1.358 | NADPH-dependent ketopantoyl lactone reductase | - |
Candida parapsilosis |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.214 | NADP+ | - |
Candida parapsilosis | |
1.1.1.214 | NADPH | - |
Candida parapsilosis | |
1.1.1.358 | NADPH | dependent on | Candida parapsilosis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.1.1.214 | evolution | the conjugated polyketone reductase C2 (CPR-C2) from Candida parapsilosis IFO 0708 belongs to the aldo-keto reductase, AKR, superfamily | Candida parapsilosis |
1.1.1.214 | additional information | catalytic tetrad in the active site of CPR-C2/NADPH | Candida parapsilosis |