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Literature summary for 2.4.2.6 extracted from

  • Del Arco, J.; Sanchez-Murcia, P.A.; Mancheno, J.M.; Gago, F.; Fernandez-Lucas, J.
    Characterization of an atypical, thermostable, organic solvent- and acid-tolerant 2'-deoxyribosyltransferase from Chroococcidiopsis thermalis (2018), Appl. Microbiol. Biotechnol., 102, 6947-6957 .
    View publication on PubMed
Search for more literature for 2.4.2.6

Application

Application Comment Organism
synthesis the enzyme is successfully employed in the enzymatic production of several therapeutic nucleosides such as didanosine, vidarabine, and cytarabine Chroococcidiopsis thermalis

Cloned(Commentary)

Cloned (Comment) Organism
gene Chro_1188, sequence comparisons, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) Chroococcidiopsis thermalis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics Chroococcidiopsis thermalis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Chroococcidiopsis thermalis CtNDT recognizes purine bases and their corresponding 2'-deoxynucleosides but is also proficient using cytosine and 2'-deoxycytidine as substrates ?
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-
additional information Chroococcidiopsis thermalis PCC 7203 CtNDT recognizes purine bases and their corresponding 2'-deoxynucleosides but is also proficient using cytosine and 2'-deoxycytidine as substrates ?
-
-

Organic Solvent Stability

Organic Solvent Comment Organism
Acetone stable at 20% v/v Chroococcidiopsis thermalis
acetonitrile stable at 20% v/v Chroococcidiopsis thermalis
chloroform loss of 30% activity at 20% v/v Chroococcidiopsis thermalis
DMSO loss of 10% activity at 20% v/v Chroococcidiopsis thermalis
Ethanol stable at 20% v/v Chroococcidiopsis thermalis
Ethyl acetate loss of 27% activity at 20% v/v Chroococcidiopsis thermalis
Ethylene glycol loss of 5% activity at 20% v/v Chroococcidiopsis thermalis
Glycerol loss of 40% activity at 20% v/v Chroococcidiopsis thermalis
isopropanol stable at 20% v/v Chroococcidiopsis thermalis
Methanol stable at 20% v/v Chroococcidiopsis thermalis
additional information CtNDT displays high activity (80-100%) in the presence of several water-miscible co-solvents in a proportion of up to 20% Chroococcidiopsis thermalis
N,N-dimethylformamide loss of 25% activity at 20% v/v Chroococcidiopsis thermalis
propylene glycol loss of 10% activity at 20% v/v Chroococcidiopsis thermalis

Organism

Organism UniProt Comment Textmining
Chroococcidiopsis thermalis K9TVX3
-
-
Chroococcidiopsis thermalis PCC 7203 K9TVX3
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, preparative gel fitration, dialysis, and ultrafiltration Chroococcidiopsis thermalis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2',3'-dideoxyadenosine + cytosine
-
 Chroococcidiopsis thermalis 2',3'-dideoxycytidine + adenine
-
 ?
2',3'-dideoxyadenosine + cytosine
-
 Chroococcidiopsis thermalis PCC 7203 2',3'-dideoxycytidine + adenine
-
 ?
2',3'-dideoxyadenosine + guanine
-
 Chroococcidiopsis thermalis 2',3'-dideoxyguanosine + adenine
-
 ?
2',3'-dideoxyadenosine + hypoxanthine
-
 Chroococcidiopsis thermalis 2',3'-dideoxyinosine + adenine
-
 ?
2',3'-dideoxyinosine + adenine
-
 Chroococcidiopsis thermalis 2',3'-dideoxyadenosine + hypoxanthine
-
 ?
2',3'-dideoxyinosine + adenine
-
 Chroococcidiopsis thermalis PCC 7203 2',3'-dideoxyadenosine + hypoxanthine
-
 ?
2',3'-dideoxyinosine + guanine
-
 Chroococcidiopsis thermalis 2',3'-dideoxyguanosine + hypoxanthine
-
 ?
2'-deoxyinosine + adenine
-
 Chroococcidiopsis thermalis 2'-deoxyadenosine + hypoxanthine
-
 ?
2'-deoxyinosine + adenine
-
 Chroococcidiopsis thermalis PCC 7203 2'-deoxyadenosine + hypoxanthine
-
 ?
2'-fluoro-2'-deoxyinosine + adenine
-
 Chroococcidiopsis thermalis 2'-fluoro-2'-deoxyadenosine + hypoxanthine
-
 ?
2'-fluoro-2'-deoxyinosine + guanine
-
 Chroococcidiopsis thermalis 2'-fluoro-2'-deoxyguanosine + hypoxanthine
-
 ?
9-beta-D-arabinofuranosyl adenine + cytosine
-
 Chroococcidiopsis thermalis ?
-
 ?
9-beta-D-arabinofuranosyl adenine + guanine
-
 Chroococcidiopsis thermalis ?
-
 ?
9-beta-D-arabinofuranosyl adenine + hypoxanthine
-
 Chroococcidiopsis thermalis ?
-
 ?
9-beta-D-arabinofuranosyl guanine + adenine
-
 Chroococcidiopsis thermalis ?
-
 ?
9-beta-D-arabinofuranosyl hypoxanthine + adenine
-
 Chroococcidiopsis thermalis ?
-
 ?
9-beta-D-arabinofuranosyl hypoxanthine + guanine
-
 Chroococcidiopsis thermalis ?
-
 ?
additional information CtNDT recognizes purine bases and their corresponding 2'-deoxynucleosides but is also proficient using cytosine and 2'-deoxycytidine as substrates Chroococcidiopsis thermalis ?
-
-
additional information CtNDT recognizes purine bases and their corresponding 2'-deoxynucleosides but is also proficient using cytosine and 2'-deoxycytidine as substrates, substrate specificity, overview. Cytidine does not react with 2',3'-dideoxyinosine, 2'-fluoro-2'-deoxyinosine, 9-beta-D-arabinofuranosyl guanine, and 9-beta-D-arabinofuranosyl hypoxanthine Chroococcidiopsis thermalis ?
-
-
additional information CtNDT recognizes purine bases and their corresponding 2'-deoxynucleosides but is also proficient using cytosine and 2'-deoxycytidine as substrates Chroococcidiopsis thermalis PCC 7203 ?
-
-
additional information CtNDT recognizes purine bases and their corresponding 2'-deoxynucleosides but is also proficient using cytosine and 2'-deoxycytidine as substrates, substrate specificity, overview. Cytidine does not react with 2',3'-dideoxyinosine, 2'-fluoro-2'-deoxyinosine, 9-beta-D-arabinofuranosyl guanine, and 9-beta-D-arabinofuranosyl hypoxanthine Chroococcidiopsis thermalis PCC 7203 ?
-
-

Subunits

Subunits Comment Organism
homotetramer
-
 Chroococcidiopsis thermalis

Synonyms

Synonyms Comment Organism
2'-deoxyribosyltransferase
-
 Chroococcidiopsis thermalis
Chro_1188
-
 Chroococcidiopsis thermalis
CtNDT
-
 Chroococcidiopsis thermalis
NDT
-
 Chroococcidiopsis thermalis
nucleoside 2-deoxyribosyltransferase
-
 Chroococcidiopsis thermalis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40 60 assay at Chroococcidiopsis thermalis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50 100 good activity and stability at Chroococcidiopsis thermalis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 8.5 assay at Chroococcidiopsis thermalis

pH Stability

pH Stability pH Stability Maximum Comment Organism
3 7 good activity and stability at Chroococcidiopsis thermalis

General Information

General Information Comment Organism
evolution a strict classification of CtNDTas either a type I or a type II NDT is not possible Chroococcidiopsis thermalis
additional information enzyme structure homology modeling, catalytic mechanism, overview. Glu88 is the catalytic residue whose carboxylate attacks the C1' of the nucleoside, is firmly positioned by the phenol of Tyr7. The carboxamide nitrogen of Asn118# and the Asp82 carboxylate recognize the O5' of the 2'-deoxyribose.Pro11 and Pro40 play a prominent role in making up the active site cavity. The heteroaromatic ring of the nucleobase is sandwiched between the phenyl rings of Phe14 and Phe41 on one side and the hydrophobic side chains of Val58 and Leu119# on the opposite side. The Met120# side chain sulfur appears close to the H1' atom, which suggests that it likely plays a role in transition state stabilization, together with the carboxylate of Asp62. The side chains of Tyr115' and possibly Asp53 can establish direct or water-bridged hydrogen bonds with the N7 of purines and the substituent on C6, be it an amino or a keto group. Asp62 of CtNDT plays a role in cytosine recognition, together with Arg43 and a protonated Asn44 Chroococcidiopsis thermalis