title: Efficient multi-enzyme-catalyzed CDP-choline production driven by an ATP donor module.
abstract: Cytidine diphosphate choline (CDP-choline) has been applied for treating acute craniocerebral injury and allowing recovery of consciousness after brain surgery. In this study, an acetate kinase (ACK)/acetyl phosphate system was used to supply ATP and combined with Escherichia coli-overexpressed CMP kinase (CMK), NDP kinase (NDK), choline phosphate cytidylyltransferase (CCT), and choline kinase (CKI) to produce CDP-choline from CMP and choline chloride. Within 1 h, 49 mM CDP-choline was produced, for a molar yield of 89.9 and 68.4 % based on CMP and choline chloride, respectively; the utilization efficiency of energy (UEE) was 79.5 %. Acetyl phosphate, sodium acetate, and CTP inhibited the reaction when the concentration exceeded 18.5, 600, and 30 mM, respectively. This inhibition could be overcome by controlling the rate of acetyl phosphate, CMP addition or using KOH instead of NaOH to regulate the pH in fed-batch transformation. After 24 h, the maximum titer was 124.1 ± 2.7 mM, the productivity was 5.1 ± 0.1 mM l-1h-1, the molar yield to CMP and choline chloride were 83.8 and 63.7 %, respectively, and the UEE was 58.2 %. This high yield and productivity of CDP-choline through biocatalysis suggest future application at the industrial scale.
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, 211816, People's Republic of China.|National Engineering Technique Research Center for Biotechnology, Nanjing, 211816, People's Republic of China.|Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), Nanjing, 211816, People's Republic of China.
authors.forename: Yingmiao
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, 211816, People's Republic of China.|National Engineering Technique Research Center for Biotechnology, Nanjing, 211816, People's Republic of China.
authors.forename: Junzhi
authors.initials: J
authors.lastname: Wang
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China.
authors.forename: Chongmao
authors.initials: C
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, 211816, People's Republic of China.|National Engineering Technique Research Center for Biotechnology, Nanjing, 211816, People's Republic of China.|Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), Nanjing, 211816, People's Republic of China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, 211816, People's Republic of China.|National Engineering Technique Research Center for Biotechnology, Nanjing, 211816, People's Republic of China.
authors.forename: Dong
authors.initials: D
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, 211816, People's Republic of China.|National Engineering Technique Research Center for Biotechnology, Nanjing, 211816, People's Republic of China.
authors.forename: Huanqing
authors.initials: H
authors.lastname: Niu
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), Nanjing, 211816, People's Republic of China. syang@sibs.ac.cn.|Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China. syang@sibs.ac.cn.
authors.forename: Sheng
authors.initials: S
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, 211816, People's Republic of China.|National Engineering Technique Research Center for Biotechnology, Nanjing, 211816, People's Republic of China.|Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), Nanjing, 211816, People's Republic of China.
authors.forename: Hanjie
authors.initials: H
authors.lastname: Ying
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D055162:Biocatalysis; D019916:Choline-Phosphate Cytidylyltransferase / Q000378:metabolism*; D003352:Corynebacterium / Q000201:enzymology* / Q000378:metabolism*; D003566:Cytidine Diphosphate Choline / Q000378:metabolism*
chemical_list: D003566:Cytidine Diphosphate Choline; D000255:Adenosine Triphosphate; D019916:Choline-Phosphate Cytidylyltransferase
keywords: Biocatalysis; CDP-choline acetate kinase/acetyl phosphate; Choline phosphate cytidylyltransferase; Corynebacterium ammoniagenes