One pot process of 2' deoxyguanylic acid catalyzed by a multi enzyme system
Abstract2' Deoxyguanylic acid (deoxyguanosine 5' monophosphate, dGMP) is a substance required by living cells that is used extensively in reagents, fine chemicals and other industrial fields. Traditionally, dGMP is separated from DNA degradation products, which is low yielding and time consuming. Herein, we investigated a novel, one pot multi enzymatic cascade reaction to produce dGMP. This reaction involved purine nucleoside phosphorylase (PNPase) and acetate kinase (ACKase) from Escherichia coli, N deoxyribosytransferase II (NDT II) from Lactobacillus delbrueckii and deoxyguanosine kinase (dGKase) from Bacillus subtilis. During the reaction, the initial guanosine substrate was cleaved into guanine and ribose 1 phosphate by PNPase. Then, deoxyguanosine (dGR) was subsequently produced from a reaction between guanine and thymidine catalysed by NDT II. Finally, the intermediate dGR was phosphorylated to dGMP by dGKase and a cytidine triphosphate (CTP) regeneration system that utilised acetyl phosphate via ACKase. A very small amount of CTP was added because CTP regeneration was efficient to transfer a phosphate group from acetyl phosphate to dGR. After 12 h of incubation, a maximal dGMP yield of up to 76% was obtained based on the addition of 5 mM guanosine and 5 mM thymidine.
Biotechnology and Bioprocess Engineering