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Literature summary for 1.1.1.358 extracted from

  • Wang, J.; Cheng, P.; Wu, Y.; Wang, A.; Liu, F.; Pei, X.
    Discovery of a new NADPH-dependent aldo-keto reductase from Candida orthopsilosis catalyzing the stereospecific synthesis of (R)-pantolactone by genome mining (2019), J. Biotechnol., 291, 26-34 .
    View publication on PubMed

Application

Application Comment Organism
synthesis the enzyme has a potential application in the asymmetric synthesis of optically active (R)-pantothenate, synthetic method of (R)-pantothenate production through the stereoselective reduction of oxopantoyl lactone (KPL) by aldo-keto reductase (AKR). (R)-Pantolactone (PL) is a key chiral building block for the synthesis of calcium (R)-pantothenate (vitamin B5), (R)-panthenol, and (R)-pantetheine, which are used as food and feed additives, as well as ingredients in pharmaceutical and cosmetic compositions Candida orthopsilosis

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) Candida orthopsilosis

Inhibitors

Inhibitors Comment Organism Structure
Ag+ complete inhibition at 1 mM Candida orthopsilosis
Co2+
-
Candida orthopsilosis
Cu2+
-
Candida orthopsilosis
EDTA
-
Candida orthopsilosis
Fe2+
-
Candida orthopsilosis
Mg2+
-
Candida orthopsilosis
Mn2+
-
Candida orthopsilosis
Pb2+ complete inhibition at 1 mM Candida orthopsilosis
Zn2+
-
Candida orthopsilosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis-Menten kintics Candida orthopsilosis
1.3
-
2-dehydropantolactone pH 7.0, 30°C, recombinant enzyme Candida orthopsilosis

Metals/Ions

Metals/Ions Comment Organism Structure
additional information Ni2+ and Ca2+ have almost no influence on the activity of CorCPR Candida orthopsilosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-dehydropantolactone + NADPH + H+ Candida orthopsilosis
-
(R)-pantolactone + NADP+
-
?
2-dehydropantolactone + NADPH + H+ Candida orthopsilosis Co 90-125
-
(R)-pantolactone + NADP+
-
?

Organism

Organism UniProt Comment Textmining
Candida orthopsilosis
-
-
-
Candida orthopsilosis Co 90-125
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis Candida orthopsilosis

Reaction

Reaction Comment Organism Reaction ID
(R)-pantolactone + NADP+ = 2-dehydropantolactone + NADPH + H+ enzyme CorCPR has a catalytic tetrad D-Y-K-H, and the detailed catalytic mechanism is clarified by molecular docking, overview Candida orthopsilosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-dehydropantolactone + NADPH + H+
-
Candida orthopsilosis (R)-pantolactone + NADP+
-
?
2-dehydropantolactone + NADPH + H+ recombinant CorCPR exhibited a high catalytic activity for the synthesis of (R)-pantolactone ((R)-PL) from 2-dehydropantolactone (KPL). The activity assay is performed using purified CorCPR and crude recombinant GDH in a single batch reaction. GDH is used to complete the enzyme-coupled NADPH regeneration system. CPR-01 form Candida orthopsilosis Co 90-125 exhibits excellent enantioselectivity (enantiomeric excess of over 99%). CorCPR exhibits the highest activity and stereoselectivity toward KPL Candida orthopsilosis (R)-pantolactone + NADP+
-
?
2-dehydropantolactone + NADPH + H+
-
Candida orthopsilosis Co 90-125 (R)-pantolactone + NADP+
-
?
2-dehydropantolactone + NADPH + H+ recombinant CorCPR exhibited a high catalytic activity for the synthesis of (R)-pantolactone ((R)-PL) from 2-dehydropantolactone (KPL). The activity assay is performed using purified CorCPR and crude recombinant GDH in a single batch reaction. GDH is used to complete the enzyme-coupled NADPH regeneration system. CPR-01 form Candida orthopsilosis Co 90-125 exhibits excellent enantioselectivity (enantiomeric excess of over 99%). CorCPR exhibits the highest activity and stereoselectivity toward KPL Candida orthopsilosis Co 90-125 (R)-pantolactone + NADP+
-
?
additional information the aldo-keto reductase (AKR) can catalyze ketopantoyl lactone to optically active (R)-pantolactone. A putative catalytic mechanism is proposed that Tyr63 acts as a general acid. No activity of recombinant CorCPR with typical AKR substrates such as p-nitrobenzaldehyde. Other alpha-oxoesters are not the optimal substrates of CorCPR due to the relatively low activities and low stereoselectivities Candida orthopsilosis ?
-
-
additional information the aldo-keto reductase (AKR) can catalyze ketopantoyl lactone to optically active (R)-pantolactone. A putative catalytic mechanism is proposed that Tyr63 acts as a general acid. No activity of recombinant CorCPR with typical AKR substrates such as p-nitrobenzaldehyde. Other alpha-oxoesters are not the optimal substrates of CorCPR due to the relatively low activities and low stereoselectivities Candida orthopsilosis Co 90-125 ?
-
-

Subunits

Subunits Comment Organism
? x * 37000, about, recombinant His6-tagged enzyme, sequence calculation and SDS-PAGE Candida orthopsilosis

Synonyms

Synonyms Comment Organism
conjugated polyketone reductase
-
Candida orthopsilosis
CorCPR
-
Candida orthopsilosis
CPR-01
-
Candida orthopsilosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
recombinant enzyme Candida orthopsilosis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 55 over 50% of maximal activity within this range Candida orthopsilosis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40 50 purified recombinant His6-tagged enzyme, pH 7.0, half-lives of the enzyme at 40, 45 and 50°C are 6.3 h, 3.7 h and 52 min, respectively Candida orthopsilosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
recombinant enzyme Candida orthopsilosis

pH Range

pH Minimum pH Maximum Comment Organism
6 8 over 40% of maximal activity within this range Candida orthopsilosis

Cofactor

Cofactor Comment Organism Structure
NADPH NADPH is embedded in the coenzyme binding groove in an extended conformation, and the nicotinamide ring positioned at the bottom of the substrate binding cavity. The adenosine 2'-phosphate group is bound to the side chains of Lys28, Ser263 and Arg267 of CorCPR Candida orthopsilosis

General Information

General Information Comment Organism
evolution enzyme CorCPR belongs to the AKR3C family of AKR superfamily, it also contains the conserved GXGT/SX motif of the AKR superfamily Candida orthopsilosis
additional information docking analysis and structure homology modeling of CorCPR, the crystal structure of CPR-C2 from Candida parapsilosis strain IFO 0708 (PDB ID 3VXG, resolution 1.7 A) is used as the model structure, overview. KPL is docked into the substrate binding pocket. The model predicts that the C3 carbonyl oxygen of KPL forms hydrogen bond with the side chains of Tyr63, and the alpha-amino group of Thr27 also forms a hydrogen bond with C2 carbonyl oxygen. The catalytic tetrads of CorCPR are Asp58, Tyr63, Lys88 and His125. Tyr63 acts as a general acid, and His125 facilitates proton donation, the phenolic hydroxy group of Tyr63 provide general acid catalytic assistance to carbonyl group of KPL, and residues Asp 58 and Tys88 are responsible for the hydrogen transfer. The stereospecificity of CorCPR is respected to the pro-R hydrogen at C4 of the nicotinamide ring and the pseudo re-side attack of the hydride on the carbonyl group. In addition, the formation of hydrogen bond between Thr27 and the C2 carbonyl oxygen of KPL plays an important role in substrate recognition Candida orthopsilosis