Abstract
Publisher Summary This chapter describes the enzyme nucleoside deoxyribosyltransferase from Lactobacillus helveticus. Nucleoside deoxydbosyltransferase (originally named trans-N-glycosidase 13, then trans-N-deoxyribosylase), catalyzes three types of transfer of the deoxyribosyl moiety according to the nature of the donor and acceptor bases. Lactobacillus helveticus extracts contain two different enzymes: nucleoside deoxyribosyltransferase-I or purine nucleoside and nucleoside deoxyribosyltransferase-II or purine (pyrimidine) nucleoside. Because the amount of these two types of enzyme varies significantly from strain to strain, it is desirable to perform the assay reactions. Hypoxanthine formed (using deoxyinosine as donor) is rapidly oxidized to uric acid by xanthine oxidase. The activity is measured at 40°. The change in absorbance is followed at 290 nm where most common bases and nucleosides have low extinction coefficients. It is important to check that the rate of the oxidation of hypoxanthine is not limiting and that no deoxyinosine hydrolase, adenine deaminase, or nucleoside phosphorylase are present.
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