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Updated: Mar 13, 2026
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A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
Published on: October 4, 2019
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Efficient multi-enzyme-catalyzed CDP-choline production driven by an ATP donor module.
Yingmiao Liu1,2,3, Junzhi Wang1,2, Chongmao Xu4
1State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, 211816, People's Republic of China.
Applied Microbiology and Biotechnology
|October 15, 2016
Summary
This study developed a biocatalytic method to produce cytidine diphosphate choline (CDP-choline) using enzymes from E. coli. The process achieved high yields and productivity, suggesting potential for industrial-scale CDP-choline synthesis.
Area of Science:
- Biotechnology
- Enzyme Engineering
- Biochemical Production
Background:
- Cytidine diphosphate choline (CDP-choline) is used therapeutically for acute craniocerebral injury and post-brain surgery consciousness recovery.
- Existing production methods may have limitations in yield and efficiency.
Purpose of the Study:
- To develop an efficient biocatalytic method for CDP-choline production.
- To optimize the enzymatic synthesis of CDP-choline using an acetate kinase/acetyl phosphate system and overexpressed enzymes.
Main Methods:
- Utilized an acetate kinase (ACK)/acetyl phosphate system for ATP regeneration.
- Employed overexpressed Escherichia coli enzymes: CMP kinase (CMK), NDP kinase (NDK), choline phosphate cytidylyltransferase (CCT), and choline kinase (CKI).
- Investigated fed-batch transformation with controlled addition rates and pH regulation to overcome product inhibition.
Main Results:
- Achieved 49 mM CDP-choline within 1 hour with high molar yields (89.9% from CMP, 68.4% from choline chloride) and 79.5% energy utilization efficiency (UEE).
- Identified and overcame inhibition by acetyl phosphate, sodium acetate, and CTP through controlled feeding and pH adjustment.
- Reached a maximum titer of 124.1 ± 2.7 mM after 24 hours with a productivity of 5.1 ± 0.1 mM/L/h and improved UEE of 58.2%.
Conclusions:
- The developed biocatalytic process demonstrates high yield and productivity for CDP-choline synthesis.
- The method shows significant potential for industrial-scale application in CDP-choline production.
- Enzymatic synthesis offers a promising alternative for manufacturing therapeutic compounds like CDP-choline.
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