Abstract |
Dephospho coenzyme A (depCoA) is the last intermediate for CoA biosynthesis, and it can be used as a transcription initiator to prepare CoA-linked RNA by in vitro transcription. However, commercially available depCoA is expensive. We hereby describe a simple and efficient enzymatic synthesis of depCoA in a single-step from commercially available and inexpensive oxidized pantethine (Ox-Pan) and ATP. A plasmid (pCoaDAa) was constructed to co-express and co-purify two enzymes pantothenate kinase (PanK/coaA) and phosphopantetheine adenylyltransferase (PPAT/ coaD). Starting from Ox-Pan and ATP, two different synthetic routes of one-pot reaction catalyzed by PanK and PPAT, followed by a simple column purification step, afforded depCoA and its oxidized dimer (Ox-depCoA) with high yields and purity. The simplicity and low cost of our method should make depCoA easily accessible to a broad scientific community, and promote research on CoA-related areas in biology and biomedicine.
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Authors | Krishna Sapkota, Faqing Huang |
Journal | Bioorganic chemistry
(Bioorg Chem)
Vol. 76
Pg. 23-27
(02 2018)
ISSN: 1090-2120 [Electronic] United States |
PMID | 29107839
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2017 Elsevier Inc. All rights reserved. |
Chemical References |
- dephosphocoenzyme A
- Pantetheine
- pantethine
- Adenosine Triphosphate
- Phosphotransferases (Alcohol Group Acceptor)
- pantothenate kinase
- Nucleotidyltransferases
- pantetheine-phosphate adenylyltransferase
- Coenzyme A
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Topics |
- Adenosine Triphosphate
(chemistry)
- Amino Acid Sequence
- Base Sequence
- Chemistry Techniques, Synthetic
(methods)
- Cloning, Molecular
(methods)
- Coenzyme A
(chemical synthesis)
- Escherichia coli
(enzymology)
- Nucleotidyltransferases
(genetics, metabolism)
- Oxidation-Reduction
- Pantetheine
(analogs & derivatives, chemistry)
- Phosphotransferases (Alcohol Group Acceptor)
(genetics, metabolism)
- Plasmids
(genetics)
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