Application | Comment | Organism |
---|---|---|
synthesis | the recombinant CduCPR from Candida dubliniensis exhibits potential application in the asymmetric synthesis of (R)-pantolactone | Candida dubliniensis |
Cloned (Comment) | Organism |
---|---|
gene CPR-C2, functional overexpression of N-terminally His6-tagged wild-type enzyme in Escherichia coli strain BL21(DE3), recombinant expression of His6-tagged enzyme mutants in Escherichia coli | Candida dubliniensis |
Protein Variants | Comment | Organism |
---|---|---|
H128A | site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type enzyme | Candida dubliniensis |
T30A | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme | Candida dubliniensis |
Y66A | site-directed mutagenesis, inactive mutant | Candida dubliniensis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ag+ | complete inhibition at 1 mM | Candida dubliniensis | |
Ca2+ | 5.4% inhibition at 1 mM | Candida dubliniensis | |
Co2+ | 24.7% inhibition at 1 mM | Candida dubliniensis | |
Cu2+ | 70.3% inhibition at 1 mM | Candida dubliniensis | |
EDTA | 14.3% inhibition at 1 mM | Candida dubliniensis | |
Fe2+ | 33.2% inhibition at 1 mM | Candida dubliniensis | |
Mg2+ | 22.4% inhibition at 1 mM | Candida dubliniensis | |
Mn2+ | 43.2% inhibition at 1 mM | Candida dubliniensis | |
Ni2+ | 11.5% inhibition at 1 mM | Candida dubliniensis | |
Pb2+ | complete inhibition at 1 mM | Candida dubliniensis | |
Zn2+ | 89.2% inhibition at 1 mM | Candida dubliniensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.484 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant wild-type enzyme | Candida dubliniensis | |
3.5 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant mutant T30A | Candida dubliniensis | |
20.4 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant mutant H128A | Candida dubliniensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-dehydropantolactone + NADPH + H+ | Candida dubliniensis | - |
(R)-pantolactone + NADP+ | - |
? | |
2-dehydropantolactone + NADPH + H+ | Candida dubliniensis ATCC MYA-646 | - |
(R)-pantolactone + NADP+ | - |
? | |
2-dehydropantolactone + NADPH + H+ | Candida dubliniensis CD36 | - |
(R)-pantolactone + NADP+ | - |
? | |
2-dehydropantolactone + NADPH + H+ | Candida dubliniensis NCPF 3949 | - |
(R)-pantolactone + NADP+ | - |
? | |
2-dehydropantolactone + NADPH + H+ | Candida dubliniensis NRRL Y-17841 | - |
(R)-pantolactone + NADP+ | - |
? | |
2-dehydropantolactone + NADPH + H+ | Candida dubliniensis CBS 7987 | - |
(R)-pantolactone + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Candida dubliniensis | B9WJQ5 | - |
- |
Candida dubliniensis ATCC MYA-646 | B9WJQ5 | - |
- |
Candida dubliniensis CBS 7987 | B9WJQ5 | - |
- |
Candida dubliniensis CD36 | B9WJQ5 | - |
- |
Candida dubliniensis NCPF 3949 | B9WJQ5 | - |
- |
Candida dubliniensis NRRL Y-17841 | B9WJQ5 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis | Candida dubliniensis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(R)-pantolactone + NADP+ = 2-dehydropantolactone + NADPH + H+ | catalytic mechanism of (R)-pantolactone synthesis, overview. The conserved catalytic triad is formed by residues Thr30, Tyr66 and His128. Tyr66 functions as a proton donor following hydrogen transfer from NADPH. Thr30 and His128 are critical residues to bind and orient 2-dehydropantolactone (KPL). The phenolic hydroxyl group of Tyr66 forms the hydrogen bond with the C5 carbonyl oxygen of 2-dehydropantolactone. The HR of NADPH could just attack the si-face of the carbonyl group, which results in the formation of (R)-pantolactone. The alpha-amino group and the hydroxyl group of Thr30 form hydrogen bonds with the C4 carbonyl oxygen of KPL, and the epsilon-amino group of Lys33 formes the hydrogen bond with the O2 of KPL. In addition, the hydrophobic interaction of His128, Val165 and Phe302 with two methyl groups of KPL might facilitate the substrate binding | Candida dubliniensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-dehydropantolactone + NADPH + H+ | - |
Candida dubliniensis | (R)-pantolactone + NADP+ | - |
? | |
2-dehydropantolactone + NADPH + H+ | - |
Candida dubliniensis ATCC MYA-646 | (R)-pantolactone + NADP+ | - |
? | |
2-dehydropantolactone + NADPH + H+ | - |
Candida dubliniensis CD36 | (R)-pantolactone + NADP+ | - |
? | |
2-dehydropantolactone + NADPH + H+ | - |
Candida dubliniensis NCPF 3949 | (R)-pantolactone + NADP+ | - |
? | |
2-dehydropantolactone + NADPH + H+ | - |
Candida dubliniensis NRRL Y-17841 | (R)-pantolactone + NADP+ | - |
? | |
2-dehydropantolactone + NADPH + H+ | - |
Candida dubliniensis CBS 7987 | (R)-pantolactone + NADP+ | - |
? | |
additional information | enzyme CduCPR shows high catalytic activity and stereoselectivity using 2-dehydropantolactone (KPL) as the substrate. In a continuous feeding reaction, 200 mM ketopantolactone is reduced to (R)-pantolactone with 98% conversion and 99% enantiomeric excess within 2.0 h by the purified recombinant enzyme | Candida dubliniensis | ? | - |
- |
|
additional information | enzyme CduCPR shows high catalytic activity and stereoselectivity using 2-dehydropantolactone (KPL) as the substrate. In a continuous feeding reaction, 200 mM ketopantolactone is reduced to (R)-pantolactone with 98% conversion and 99% enantiomeric excess within 2.0 h by the purified recombinant enzyme | Candida dubliniensis ATCC MYA-646 | ? | - |
- |
|
additional information | enzyme CduCPR shows high catalytic activity and stereoselectivity using 2-dehydropantolactone (KPL) as the substrate. In a continuous feeding reaction, 200 mM ketopantolactone is reduced to (R)-pantolactone with 98% conversion and 99% enantiomeric excess within 2.0 h by the purified recombinant enzyme | Candida dubliniensis CD36 | ? | - |
- |
|
additional information | enzyme CduCPR shows high catalytic activity and stereoselectivity using 2-dehydropantolactone (KPL) as the substrate. In a continuous feeding reaction, 200 mM ketopantolactone is reduced to (R)-pantolactone with 98% conversion and 99% enantiomeric excess within 2.0 h by the purified recombinant enzyme | Candida dubliniensis NCPF 3949 | ? | - |
- |
|
additional information | enzyme CduCPR shows high catalytic activity and stereoselectivity using 2-dehydropantolactone (KPL) as the substrate. In a continuous feeding reaction, 200 mM ketopantolactone is reduced to (R)-pantolactone with 98% conversion and 99% enantiomeric excess within 2.0 h by the purified recombinant enzyme | Candida dubliniensis NRRL Y-17841 | ? | - |
- |
|
additional information | enzyme CduCPR shows high catalytic activity and stereoselectivity using 2-dehydropantolactone (KPL) as the substrate. In a continuous feeding reaction, 200 mM ketopantolactone is reduced to (R)-pantolactone with 98% conversion and 99% enantiomeric excess within 2.0 h by the purified recombinant enzyme | Candida dubliniensis CBS 7987 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
CduCPR | - |
Candida dubliniensis |
CPR | - |
Candida dubliniensis |
CPR-C2 | - |
Candida dubliniensis |
NADPH-dependent conjugated polyketone reductase | - |
Candida dubliniensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 45 | recombinant enzyme | Candida dubliniensis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 55 | over 40% of maximal activity within this range | Candida dubliniensis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
purified recombinant His-tagged enzyme, pH 7.0, 7 h, about 90% activity remaining | Candida dubliniensis |
40 | - |
purified recombinant His-tagged enzyme, pH 7.0, 7 h, about 50% activity remaining | Candida dubliniensis |
50 | - |
purified recombinant His-tagged enzyme, pH 7.0, 7 h, about 10% activity remaining | Candida dubliniensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
6.01 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant mutant H128A | Candida dubliniensis | |
15.5 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant mutant T30A | Candida dubliniensis | |
59 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant wild-type enzyme | Candida dubliniensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
recombinant enzyme | Candida dubliniensis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5.5 | 8.5 | over 40% of maximal activity within this range | Candida dubliniensis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Candida dubliniensis |
General Information | Comment | Organism |
---|---|---|
evolution | enzyme CduCPR belongs to the aldo-keto reductase superfamily | Candida dubliniensis |
additional information | homology modeled structure of CduCPR and molecular docking analysis, overview | Candida dubliniensis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.295 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant mutant H128A | Candida dubliniensis | |
4.43 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant mutant T30A | Candida dubliniensis | |
121.9 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant wild-type enzyme | Candida dubliniensis |