文献类型：期刊文献

英文题名：One-pot process of 2'-deoxyguanylic acid catalyzed by a multi-enzyme system

作者：Li, Yanyu[1];Ding, Qingbao[1];Ou, Ling[1];Qian, Yahui[1];Zhang, Jiao[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2015

卷号：20

期号：1

起止页码：37

外文期刊名：BIOTECHNOLOGY AND BIOPROCESS ENGINEERING

收录：;EI(收录号：20151400699201);WOS:【SCI-EXPANDED(收录号:WOS:000351466100005)】；

语种：英文

外文关键词：2 '-deoxyguanylic acid; N-deoxyribosyltransferase; purine nucleoside phosphorylase; deoxyguanosine kinase; acetate kinase

摘要：2'-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.