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Efficient one-pot enzymatic synthesis of dephospho coenzyme A.

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.
AuthorsKrishna Sapkota, Faqing Huang
JournalBioorganic chemistry (Bioorg Chem) Vol. 76 Pg. 23-27 (02 2018) ISSN: 1090-2120 [Electronic] United States
PMID29107839 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright © 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
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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