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Literature summary for 1.1.1.168 extracted from

  • Cheng, P.; Wang, J.; Wu, Y.; Jiang, X.; Pei, X.; Su, W.
    Recombinant expression and molecular insights into the catalytic mechanism of an NADPH-dependent conjugated polyketone reductase for the asymmetric synthesis of (R)-pantolactone (2019), Enzyme Microb. Technol., 126, 77-85 .
    View publication on PubMed
Search for more literature for 1.1.1.168

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Candida dubliniensis

Protein Variants

Protein Variants Comment Organism
H128A the mutant shows strongly reduced activity compared to the wild type enzyme Candida dubliniensis
T30A the mutant shows reduced activity compared to the wild type enzyme Candida dubliniensis
Y66A inactive Candida dubliniensis

Inhibitors

Inhibitors Comment Organism Structure
Ag+ complete inhibition at 1 mM Candida dubliniensis
Ca2+ 94.6% residual activity at 1 mM Candida dubliniensis
Co2+ 75.3% residual activity at 1 mM Candida dubliniensis
Cu2+ 29.7% residual activity at 1 mM Candida dubliniensis
EDTA 85.7% residual activity at 1 mM Candida dubliniensis
Fe2+ 66.8% residual activity at 1 mM Candida dubliniensis
Mg2+ 77.6% residual activity at 1 mM Candida dubliniensis
Mn2+ 56.8% residual activity at 1 mM Candida dubliniensis
Ni2+ 88.5% residual activity at 1 mM Candida dubliniensis
Pb2+ complete inhibition at 1 mM Candida dubliniensis
Zn2+ 10.8% residual activity at 1 mM Candida dubliniensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.484
-
 2-dehydropantolactone wild type enzyme, at pH 7.0 and 30°C Candida dubliniensis
3.5
-
 2-dehydropantolactone mutant enzyme T30A, at pH 7.0 and 30°C Candida dubliniensis
20.4
-
 2-dehydropantolactone mutant enzyme H128A, at pH 7.0 and 30°C Candida dubliniensis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-dehydropantolactone + NADPH + H+ Candida dubliniensis 98% conversion and 99% enantiomeric excess within 2 h (R)-pantolactone + NADP+
-
 ?
2-dehydropantolactone + NADPH + H+ Candida dubliniensis ATCC MYA-646 98% conversion and 99% enantiomeric excess within 2 h (R)-pantolactone + NADP+
-
 ?
2-dehydropantolactone + NADPH + H+ Candida dubliniensis CD36 98% conversion and 99% enantiomeric excess within 2 h (R)-pantolactone + NADP+
-
 ?
2-dehydropantolactone + NADPH + H+ Candida dubliniensis NCPF 3949 98% conversion and 99% enantiomeric excess within 2 h (R)-pantolactone + NADP+
-
 ?
2-dehydropantolactone + NADPH + H+ Candida dubliniensis NRRL Y-17841 98% conversion and 99% enantiomeric excess within 2 h (R)-pantolactone + NADP+
-
 ?
2-dehydropantolactone + NADPH + H+ Candida dubliniensis CBS 7987 98% conversion and 99% enantiomeric excess within 2 h (R)-pantolactone + NADP+
-
 ?

Organism

Organism UniProt Comment Textmining
Candida dubliniensis B9WJQ5
-
-
Candida dubliniensis CD36 B9WJQ5
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Candida dubliniensis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
10.3
-
 mutant enzyme H128A, at pH 7.0 and 30°C Candida dubliniensis
26.5
-
 mutant enzyme T30A, at pH 7.0 and 30°C Candida dubliniensis
49.1
-
 wild type enzyme, at pH 7.0 and 30°C Candida dubliniensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-dehydropantolactone + NADPH + H+ 98% conversion and 99% enantiomeric excess within 2 h Candida dubliniensis (R)-pantolactone + NADP+
-
 ?
2-dehydropantolactone + NADPH + H+ 98% conversion and 99% enantiomeric excess within 2 h Candida dubliniensis ATCC MYA-646 (R)-pantolactone + NADP+
-
 ?
2-dehydropantolactone + NADPH + H+ 98% conversion and 99% enantiomeric excess within 2 h Candida dubliniensis CD36 (R)-pantolactone + NADP+
-
 ?
2-dehydropantolactone + NADPH + H+ 98% conversion and 99% enantiomeric excess within 2 h Candida dubliniensis NCPF 3949 (R)-pantolactone + NADP+
-
 ?
2-dehydropantolactone + NADPH + H+ 98% conversion and 99% enantiomeric excess within 2 h Candida dubliniensis NRRL Y-17841 (R)-pantolactone + NADP+
-
 ?
2-dehydropantolactone + NADPH + H+ 98% conversion and 99% enantiomeric excess within 2 h Candida dubliniensis CBS 7987 (R)-pantolactone + NADP+
-
 ?

Subunits

Subunits Comment Organism
? x * 37000, SDS-PAGE Candida dubliniensis

Synonyms

Synonyms Comment Organism
CPR
-
 Candida dubliniensis
NADPH-dependent conjugated polyketone reductase
-
 Candida dubliniensis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
 Candida dubliniensis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
35 55 between 35 and 55°C, the enzyme activity remains above 70% of the maximal activity Candida dubliniensis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30 50 the residual activity of recombinant enzyme maintains above 90% of the initial activity after 7 h at 30°C. When the enzyme is incubated at 50°C, above 30% of the initial activity still retains after 2 h. The recombinant enzyme exhibits adequate thermal stability, and its half-lives are 45.6, 8.6 and 1.7 h at 30, 40 and 50°C, respectively Candida dubliniensis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.01
-
 2-dehydropantolactone mutant enzyme H128A, at pH 7.0 and 30°C Candida dubliniensis
15.5
-
 2-dehydropantolactone mutant enzyme T30A, at pH 7.0 and 30°C Candida dubliniensis
59
-
 2-dehydropantolactone wild type enzyme, at pH 7.0 and 30°C Candida dubliniensis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
 Candida dubliniensis

pH Stability

pH Stability pH Stability Maximum Comment Organism
6.5 7 the half-lives of the enzyme are 12.8 and 8.9 h at pH 6.5 and pH 7.0, respectively Candida dubliniensis

Cofactor

Cofactor Comment Organism Structure
NADP+
-
 Candida dubliniensis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.295
-
 2-dehydropantolactone mutant enzyme H128A, at pH 7.0 and 30°C Candida dubliniensis
4.43
-
 2-dehydropantolactone mutant enzyme T30A, at pH 7.0 and 30°C Candida dubliniensis
122
-
 2-dehydropantolactone wild type enzyme, at pH 7.0 and 30°C Candida dubliniensis