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Literature summary for 1.1.1.214 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
Search for more literature for 1.1.1.214

Cloned(Commentary)

Cloned (Comment) Organism
gene cpr-c2, recombinant expression of His-tagged enzyme in Escherichia coli strain Rosetta (DE3) Candida parapsilosis

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants

Protein Variants Comment Organism
D58A site-directed mutagenesis, the mutant shows 4.82% of wild-type activity Candida parapsilosis
F299A site-directed mutagenesis, the mutant shows 19.1% of wild-type activity Candida parapsilosis
F300A site-directed mutagenesis, the mutant shows 17.8% of wild-type activity Candida parapsilosis
H125A site-directed mutagenesis, the mutant shows 1.5% of wild-type activity Candida parapsilosis
K264A site-directed mutagenesis, the mutant shows 65.7% of wild-type activity Candida parapsilosis
K28A site-directed mutagenesis, the mutant shows 71.1% of wild-type activity Candida parapsilosis
K30A site-directed mutagenesis, the mutant shows 55.1% of wild-type activity Candida parapsilosis
K88A site-directed mutagenesis, inactive mutant Candida parapsilosis
R267A site-directed mutagenesis, the mutant shows 8.43% of wild-type activity Candida parapsilosis
T27A site-directed mutagenesis, the mutant shows 5.75% of wild-type activity Candida parapsilosis
Y63A site-directed mutagenesis, the mutant shows 0.17% of wild-type activity Candida parapsilosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(R)-pantolactone + NADP+ Candida parapsilosis
-
 2-dehydropantolactone + NADPH + H+
-
 r
(R)-pantolactone + NADP+ Candida parapsilosis IFO 0708
-
 2-dehydropantolactone + NADPH + H+
-
 r
2-dehydropantolactone + NADPH + H+ Candida parapsilosis the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner (R)-pantolactone + NADP+
-
 r
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

Organism

Organism UniProt Comment Textmining
Candida parapsilosis Q76L36
-
-
Candida parapsilosis IFO 0708 Q76L36
-
-

Purification (Commentary)

Purification (Comment) Organism
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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
23.6
-
 purified recombinant detagged wild-type enzyme, pH and temperature not specified in the publication Candida parapsilosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-pantolactone + NADP+
-
 Candida parapsilosis 2-dehydropantolactone + NADPH + H+
-
 r
(R)-pantolactone + NADP+
-
 Candida parapsilosis IFO 0708 2-dehydropantolactone + NADPH + H+
-
 r
2-dehydropantolactone + NADPH + H+ the enzyme reduces ketopantoyl lactone to D-pantoyl lactone in a strictly stereospecific manner Candida parapsilosis (R)-pantolactone + NADP+
-
 r
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
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 ?
-
 ?
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 ?
-
 ?

Synonyms

Synonyms Comment Organism
conjugated polyketone reductase C2
-
 Candida parapsilosis
CPR-C2
-
 Candida parapsilosis
NADPH-dependent ketopantoyl lactone reductase
-
 Candida parapsilosis
nicotinamide adenine dinucleotide phosphate-dependent ketopantoyl lactone reductase
-
 Candida parapsilosis

Cofactor

Cofactor Comment Organism Structure
NADP+
-
 Candida parapsilosis
NADPH
-
 Candida parapsilosis

General Information

General Information Comment Organism
evolution the conjugated polyketone reductase C2 (CPR-C2) from Candida parapsilosis IFO 0708 belongs to the aldo-keto reductase, AKR, superfamily Candida parapsilosis
additional information catalytic tetrad in the active site of CPR-C2/NADPH Candida parapsilosis