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Enzyme Activity Measurement for Nucleoside Deoxyribosyltransferase Using Chromatographic Assays

Creative Enzymes is the most experienced biotech company specialized in enzyme assays. Equipped with the latest instruments, the up-to-date technologies, and an outstanding expert team, Creative Enzymes is fully prepared to offer the reliable assay for transferase enzymes, including nucleoside deoxyribosyltransferase.

Nucleoside deoxyribosyltransferases (EC 2.4.2.6) catalyze the exchange between the purine or pyrimidine base of 2’-deoxyribonucleosides and free pyrimidine or purine bases. These enzymes provide an alternative to the nucleoside phosphorylases that catalyze the same process. These enzymes are specific for 2’-deoxyribonucleosides, highly regioselective, as well as stereoselective for that β-anomers are exclusively formed. Nucleoside deoxyribosyltransferases were initially described for lactobacilli and have also been found in certain species of Streptococcus and in parasitic unicellular eukaryotic organisms such as Crithidia luciliae, Trypanosoma brucei, and Borrelia burgdorferi.

Figure 1: The reaction catalyzed by nucleoside deoxyribosyltransferases. E, enzyme; B1 and B2, purine or pyrimidine. Figure 1: The reaction catalyzed by nucleoside deoxyribosyltransferases. E, enzyme; B1 and B2, purine or pyrimidine.
Reference: Fernándezlucas J, et al. Applied & Environmental Microbiology, 2010, 76(5):1462-70.

Two types of nucleoside deoxyribosyltransferases have been described in lactobacilli: type I is purine deoxyribosyltransferase, specific for the transfer of 2’-deoxyribose between two purines; type II is nucleoside 2’-deoxyribosyltransferase, which catalyzes the transfer of deoxyribose between either purines or pyrimidines. Both enzymes follow a ping-pong bi-bi mechanism by formation of a covalent deoxyriboxyl enzyme intermediate. Therefore, these enzymes can be used to carry out a ‘one-pot’ synthesis of 2’-deoxyribosylnucleosides. This biotransformation is a promising process because the reactions take place under very mild conditions and offer environmentally clean chemical technologies.

The chromatographic assay was demonstrated to be the most reliable method for measuring the activity of nucleoside deoxyribosyltransferases. Creative Enzymes presents the most advanced chromatographic techniques to measure the enzymatic activity accurately. For instance, the production of deoxyadenosine is analyzed using high performance liquid chromatography at an absorbance of 254nm, or the formed deoxycytidine is separated by reverse-phase high-performance liquid chromatography and measured at an absorbance of 260nm. Creative Enzymes promises that it will deliver the products in the shortest span of time from the date of order placement. The prompt service, best customer care, and dedicated resources have made Creative Enzymes the most preferred supplier to all clients.

Figure 2: The crystal structure of nucleoside deoxyribosyltransferase from Lactobacillus leichmannii. Figure 2: The crystal structure of nucleoside deoxyribosyltransferase from Lactobacillus leichmannii.
PDB: 1F8X

Our Products Cannot Be Used As Medicines Directly For Personal Use.