文献类型：期刊文献

英文题名：In vitro biosynthesis of ATP from adenosine and polyphosphate

作者：Sun, Chuanqi[1];Li, Zonglin[1];Ning, Xiao[1];Xu, Wentian[1];Li, Zhimin[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg Technol, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2021

卷号：8

期号：1

外文期刊名：BIORESOURCES AND BIOPROCESSING

收录：;EI(收录号：20224513081498);WOS:【SCI-EXPANDED(收录号:WOS:000723640200001)】；

基金：This study was supported by the National Natural Science Foundation of China (Grant No. 32171478), the Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project (Grant No. TSBICIP-KJGG-009), and the China National Key Research and Development Program (Grant No. 2019YFA0904300).

语种：英文

外文关键词：Adenosine triphosphate; In vitro; Multi-enzymatic cascade catalysis; Polyphosphate kinase; Adenosine kinase

摘要：Adenosine triphosphate (ATP) acts as a crucial energy currency in vivo, and it is a widely used energy and/or phosphate donor for enzyme-catalyzed reactions in vitro. In this study, we established an in vitro multi-enzyme cascade system for ATP production. Using adenosine and inorganic polyphosphate (polyP) as key substrates, we combined adenosine kinase and two functionally distinct polyphosphate kinases (PPKs) in a one-pot reaction to achieve chain-like ATP regeneration and production. Several sources of PPK were screened and characterized, and two suitable PPKs were selected to achieve high rates of ATP production. Among these, Sulfurovum lithotrophicum PPK (SlPPK) exhibited excellent activity over a wide pH range (pH 4.0-9.0) and synthesized ATP from ADP using short-chain polyP. Furthermore, it had a half-life > 155.6 h at 45 degrees C. After optimizing the reaction conditions, we finally carried out the coupling-catalyzed reaction with different initial adenosine concentrations of 10, 20, and 30 mM. The highest yields of ATP were 76.0, 70.5, and 61.3%, respectively.