Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Qin, H.M.; Yamamura, A.; Miyakawa, T.; Kataoka, M.; Nagai, T.; Kitamura, N.; Urano, N.; Maruoka, S.; Ohtsuka, J.; Nagata, K.; Shimizu, S.; Tanokura, M.
    Structure of conjugated polyketone reductase from Candida parapsilosis IFO 0708 reveals conformational changes for substrate recognition upon NADPH binding (2013), Appl. Microbiol. Biotechnol., 98, 243-249.
    View publication on PubMed

Cloned(Commentary)

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

Crystallization (Commentary)

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 Tris–HCl, 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 Tris–HCl (pH 8.1) and 23% (w/v) polyethylene glycol 3350, at 20°C Candida parapsilosis

Protein Variants

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

Natural Substrates/ Products (Substrates)

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

Organism

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
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Synonyms

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

Cofactor

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

General Information

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