文献类型：期刊文献

英文题名：Biosynthesis of (deoxy)guanosine-5′-triphosphate by GMP kinase and acetate kinase fixed on the surface of E. coli

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

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

年份：2019

卷号：122

起止页码：82

外文期刊名：ENZYME AND MICROBIAL TECHNOLOGY

收录：;EI(收录号：20190106335162);WOS:【SCI-EXPANDED(收录号:WOS:000457511400009)】；

基金：This study was supported by Shanghai Sinobest Biotechnology Co., Ltd.

语种：英文

外文关键词：(Deoxy)guanosine-5 '-triphosphate; GYP kinase; Cell surface display; Ice nucleation protein; ATP regeneration

摘要：(Deoxy)guanosine-5'-triphosphate (5'-(d)GTP), the precursor for synthesizing DNA or RNA in vivo, is an important raw material for various modem biotechnologies based on PCR. In this study, we investigated the application of whole-cell catalysts constructed by bacterial cell surface display in biosynthetic reactions of 5'(d)GTP from (deoxy)guanosine-5'-monophosphate (5'-(d)GMP). By N-terminal or N. and C-terminal fusion of the ice nucleation protein, we successfully displayed the GMP kinase of Lactobacillus bulgaricus and the acetate kinase of E. coil on the surface of E. coil cells. A large amount of soluble target protein was obtained upon induction with 0.2 mM IPTG at 25 degrees C for 30 h. The conversion of dGMP was up to 91% when catalysed by the surface-displayed enzymes at 37 degrees C for 4 h. Up to 95% of the GMP was converted after 3 h of reaction. The stability of the whole-cell catalyst at 37 degrees C was very good. The enzyme activity was maintained above 50% after 9 rounds of recovery. Our research showed that only one-twentieth of the initial substrate concentration of added ATP was sufficient to meet the reaction requirements.