文献类型：期刊文献

英文题名：Enzymatic Synthesis of Nucleoside Triphosphates and Deoxynucleoside Triphosphates by Surface-Displayed Kinases

作者：Ding, Yi[1];Ou, Ling[1];Ding, Qingbao[2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Rutgers State Univ, New Jersey Med Sch, Dept Microbiol Biochem & Mol Genet, Newark, NJ 07103 USA

年份：2020

卷号：190

期号：4

起止页码：1271

外文期刊名：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY

收录：;EI(收录号：20194707725861);WOS:【SCI-EXPANDED(收录号:WOS:000497190500001)】；

语种：英文

外文关键词：Nucleoside triphosphates; Deoxynucleoside triphosphates; Nucleotide kinase; Acetate kinase; Ice nucleation protein; Surface display; ATP regeneration

摘要：Nucleoside triphosphates and deoxynucleoside triphosphates are important biochemical molecules. In this study, recombinant Escherichia coli that could display nucleotide kinases (INP-N-NMKases) and acetate kinase (INP-N-ACKase) on the cell surface were constructed by fusing an enzyme (NMKase/ACKase) to the N-terminus of ice nucleation protein (INP-N). By using intact recombinant bacteria cells as a catalyst coupled with an ACKase-catalyzed adenosine-5 '-triphosphate (ATP) regeneration system, nucleoside triphosphates (NTPs) and deoxynucleoside triphosphates (dNTPs) could be synthesized efficiently. In a reaction system with 5 mmol/l substrate, the conversion rates of cytidine-5 '-triphosphate (CTP) and deoxycytidine-5 '-triphosphate (dCTP) were 96% and 93%, respectively, the conversion rate of ATP and deoxyadenosine-5 '-triphosphate (dATP) was 96%, the conversion rate of deoxythymidine-5 '-triphosphate (dTTP) was 91%, and the conversion rate of uridine-5 '-triphosphate (UTP) was 80%. There was no obvious degradation. At 37 degrees C, the stability of the surface-displayed fusion protein, especially in the presence of the substrate, was significantly improved. Each whole cell could be reused more than 8 times.