One-pot process of 2′-deoxyguanylic acid catalyzed by a multi-enzyme system

• Published in: Biotechnology and bioprocess engineering Vol. 20; no. 1; pp. 37 - 43
• Main Authors: Li, Yanyu, Ding, Qingbao, Ou, Ling, Qian, Yahui, Zhang, Jiao
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer-Verlag 01-02-2015; The Korean Society for Biotechnology and Bioengineering; Springer Nature B.V; 한국생물공학회
• Subjects: acetate kinase; Acids; Analysis; Aqueous solutions; Bacillus subtilis; bioprocess engineering; Biotechnology; Chemicals; Chemistry; Chemistry and Materials Science; Cloning; cytidine triphosphate; Degradation products; Deoxyribonucleic acid; DNA; E coli; Enzymes; Escherichia coli; Genetic engineering; guanine; guanosine; Industrial and Production Engineering; Kinases; Laboratories; Lactobacillus delbrueckii; phosphorylase; Plasmids; Reagents; Research Paper; Studies; thymidine; 생물공학; acetate kinase; purine nucleoside phosphorylase; deoxyguanosine kinase; 2′-deoxyguanylic acid; N-deoxyribosyltransferase
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-014-0392-y

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.