Cloned (Comment) | Organism |
---|---|
gene CPR-C2, functional overexpression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Candida dubliniensis |
Protein Variants | Comment | Organism |
---|---|---|
additional information | an effective biocatalytic cascade was developed by encapsulating a conjugated polyketone reductase (CPR), glucose dehydrogenase (GDH) and the coenzyme NADP+ in organic-inorganic hybrid nanoflowers (hNFs) for the asymmetric reduction of oxopantolactone (KPL) to synthesize (R)-(-)-pantolactone ((R)-PL). When CduCPR and TgGDH are confined in the sodium alginate (SA)-coated hNFs [CduCPR/TgGDH-Ca3(PO4)2], both of them have excellent reusability and high stability. The SA-coated hNF reactor successfully catalyzes the asymmetric synthesis of (R)-PL, which exhibits satisfactory stereoselectivity and promotes reusability in repeated batches. The SA coating functioned to competently immobilize the coenzyme NADP+ on the CduCPR/TgGDH-Ca3(PO4)2 hNFs, which maintains the maximum bioactivity to synthesize (R)-PL without exogenous coenzyme addition. Method development, optimization, and evaluation, overview | Candida dubliniensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.484 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant free enzyme | Candida dubliniensis | |
0.886 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant immobilized enzyme | 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 |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
22 | - |
purified immobilized enzyme CduCPR, pH 7.0, 30°C | Candida dubliniensis |
49.1 | - |
purified free enzyme CduCPR, pH 7.0, 30°C | 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 | product identification and quantification by gas chromatography. Optimization of the reaction conditions for the synthesis of (R)-(-)-pantolactone | Candida dubliniensis | ? | - |
- |
|
additional information | product identification and quantification by gas chromatography. Optimization of the reaction conditions for the synthesis of (R)-(-)-pantolactone | Candida dubliniensis ATCC MYA-646 | ? | - |
- |
|
additional information | product identification and quantification by gas chromatography. Optimization of the reaction conditions for the synthesis of (R)-(-)-pantolactone | Candida dubliniensis CD36 | ? | - |
- |
|
additional information | product identification and quantification by gas chromatography. Optimization of the reaction conditions for the synthesis of (R)-(-)-pantolactone | Candida dubliniensis NCPF 3949 | ? | - |
- |
|
additional information | product identification and quantification by gas chromatography. Optimization of the reaction conditions for the synthesis of (R)-(-)-pantolactone | Candida dubliniensis NRRL Y-17841 | ? | - |
- |
|
additional information | product identification and quantification by gas chromatography. Optimization of the reaction conditions for the synthesis of (R)-(-)-pantolactone | Candida dubliniensis CBS 7987 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
CduCPR | - |
Candida dubliniensis |
conjugated polyketone reductase | - |
Candida dubliniensis |
CPR | - |
Candida dubliniensis |
CPR-C2 | - |
Candida dubliniensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 45 | recombinant enzyme | Candida dubliniensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
13.6 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant immobilized enzyme | Candida dubliniensis | |
59 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant free enzyme | Candida dubliniensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
recombinant enzyme | Candida dubliniensis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Candida dubliniensis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
15.35 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant immobilized enzyme | Candida dubliniensis | |
121.9 | - |
2-dehydropantolactone | pH 7.0, 30°C, recombinant free enzyme | Candida dubliniensis |