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Literature summary for 2.4.2.6 extracted from

  • Mendez, M.B.; Rivero, C.W.; Lopez-Gallego, F.; Guisan, J.M.; Trelles, J.A.
    Development of a high efficient biocatalyst by oriented covalent immobilization of a novel recombinant 2-N-deoxyribosyltransferase from Lactobacillus animalis (2018), J. Biotechnol., 270, 39-43 .
    View publication on PubMed
Search for more literature for 2.4.2.6

Application

Application Comment Organism
synthesis the enzyme is used for a green bioprocess by employing an environmentally friendly methodology to produce floxuridine (5-fluoro-2'-deoxyuridine), a compound with proven anti-tumor activity. The enzyme as enzymatic biocatalyst meets the requirements of high activity, stability, and short reaction times needed for low-cost production in a future preparative application Ligilactobacillus animalis

Cloned(Commentary)

Cloned (Comment) Organism
gene ndt, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Ligilactobacillus animalis

Protein Variants

Protein Variants Comment Organism
additional information immobilization of the purified recombinant enzyme for use as a biocatalyst. The biocatalyst stability is significantly enhanced by multipoint covalent immobilization using a hetero-functional support activated with nickel-chelates and glyoxyl groups to promote the irreversible multivalent attachment (Ni2+-Gx) through three reaction steps 1. affinity binding between the his-tag of the enzyme and the metals on the support, 2. covalent multi-valent attachment at alkaline pH by lysine residues attachment to the glyoxyl groups of the carrier and 3. mild reduction in order to turn the reversible Schiff's base formed into irreversible (secondary amino) bonds and transform the remaining aldehyde groups into inert hydroxyl moieties. The immobilized enzyme can be reused for more than 300 h and stored during almost 3 months without activity loss. The derivative (Ni2+-30 Gx-LaNDT) is able to biosynthesize 88 mg floxuridine/g biocatalyst after 1h of reaction. The floxuridine yield is not affected significantly when the biocatalyst obtained by affinity immobilization (Ni2+-LaNDT) is assayed, but a decrease of around 50% of enzymatic activity is observed when Ni2+-Gx-LaNDT derivative is used. Structure modeling of LaNDT is used to analyze the effect of immobilization on some of the physicochemical properties of the immobilized NDT, overview Ligilactobacillus animalis

Localization

Localization Comment Organism GeneOntology No. Textmining
soluble
-
 Ligilactobacillus animalis
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-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-deoxy-D-ribosyl-base1 + base2 Ligilactobacillus animalis
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 2-deoxy-D-ribosyl-base2 + base1
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 ?
2-deoxy-D-ribosyl-base1 + base2 Ligilactobacillus animalis ATCC 35046
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 2-deoxy-D-ribosyl-base2 + base1
-
 ?

Organism

Organism UniProt Comment Textmining
Ligilactobacillus animalis
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-
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Ligilactobacillus animalis ATCC 35046
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-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, method optimization for LaNDT Ligilactobacillus animalis

Storage Stability

Storage Stability Organism
4°C, the recombinant immobilized enzyme can be stored during almost 3 months without activity loss. When storage stability at 4°C s evaluated, Ni2+-Gx-LaNDT retains full activity for more than 78 days, whereas Ni2+-LaNDT is inactive after 65 days Ligilactobacillus animalis
50°C, Ni2+-Gx-LaNDT biocatalyst is stable after 250 h of incubation at 50°C, a condition in which the ionic biocatalyst released part of its enzymatic charge during the first 6 h of incubation Ligilactobacillus animalis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2'-deoxythymidine + 5-fluorouracil
-
 Ligilactobacillus animalis 5-fluoro-2'-deoxyuridine + thymine
-
 ?
2'-deoxythymidine + 5-fluorouracil
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 Ligilactobacillus animalis ATCC 35046 5-fluoro-2'-deoxyuridine + thymine
-
 ?
2-deoxy-D-ribosyl-base1 + base2
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 Ligilactobacillus animalis 2-deoxy-D-ribosyl-base2 + base1
-
 ?
2-deoxy-D-ribosyl-base1 + base2
-
 Ligilactobacillus animalis ATCC 35046 2-deoxy-D-ribosyl-base2 + base1
-
 ?
additional information the recombinant enzyme exhibits hydrolytic activity for both purine and pyrimidine nucleosides Ligilactobacillus animalis ?
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-
additional information the recombinant enzyme exhibits hydrolytic activity for both purine and pyrimidine nucleosides Ligilactobacillus animalis ATCC 35046 ?
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-

Subunits

Subunits Comment Organism
? x * 27000, recombinant soluble enzyme, SDS-PAGE Ligilactobacillus animalis

Synonyms

Synonyms Comment Organism
2'-N-deoxyribosyltransferase
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 Ligilactobacillus animalis
LaNDT
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 Ligilactobacillus animalis
NDT
-
 Ligilactobacillus animalis

General Information

General Information Comment Organism
evolution enzyme LaNDT can be classified as 98 NDT type II Ligilactobacillus animalis
additional information structure modeling of LaNDT Ligilactobacillus animalis