Abstract
The production of deoxyribonucleoside triphosphates (dNTPs) from deoxyribonucleoside monophosphates (dNMPs) was investigated using dried baker's yeast in the presence of a high concentration of inorganic phosphate. The amount of dATP formed by 150mg/ml of yeast cells was 127 mM, with a 84% yield of the substrate, dAMP, after 18 hr incubation at 28°C with 150 mM dAMP, 800mM glucose, 800 mM potassium phosphate buffer (pH 7.0) and 10mM MgSO4. The amount of dGTP or dCTP formed was in the range of 26-34mM, from 100 mM of either dGMP or dCMP, respectively, after 8 hr incubation, but dTTP was not formed during the incubation period.
In a reaction mixture containing a dNMP-mixture, which was prepared through the hydrolysis of salmon testes DNA or herring sperm DNA (100mg/ml) with Nuclease P1, 32-49 HIM dNTP was obtained.
