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Efficient multi-enzyme-catalyzed CDP-choline production driven by an ATP donor module

  • Biotechnological products and process engineering
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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−1 h−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.

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Authors and Affiliations

  1. 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

    Yingmiao Liu, Junzhi Wang, Yong Chen, Dong Liu, Huanqing Niu & Hanjie Ying

  2. National Engineering Technique Research Center for Biotechnology, Nanjing, 211816, People’s Republic of China

    Yingmiao Liu, Junzhi Wang, Yong Chen, Dong Liu, Huanqing Niu & Hanjie Ying

  3. Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), Nanjing, 211816, People’s Republic of China

    Yingmiao Liu, Yong Chen, Sheng Yang & Hanjie Ying

  4. Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China

    Chongmao Xu, Junjie Yang, Yu Jiang & Sheng Yang

Authors
  1. Yingmiao Liu
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  2. Junzhi Wang
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  3. Chongmao Xu
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  4. Yong Chen
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  5. Junjie Yang
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  6. Dong Liu
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  7. Huanqing Niu
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  8. Yu Jiang
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  9. Sheng Yang
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  10. Hanjie Ying
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Correspondence to Sheng Yang.

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Liu, Y., Wang, J., Xu, C. et al. Efficient multi-enzyme-catalyzed CDP-choline production driven by an ATP donor module. Appl Microbiol Biotechnol 101, 1409–1417 (2017). https://doi.org/10.1007/s00253-016-7874-0

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  • DOI: https://doi.org/10.1007/s00253-016-7874-0

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