Efficient One-Pot Enzymatic Synthesis of Dephospho Coenzyme A
Document Type
Article
Publication Date
2-2018
Department
Chemistry and Biochemistry
School
Mathematics and Natural Sciences
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.
Publication Title
Bioorganic Chemistry
Volume
76
First Page
23
Last Page
27
Recommended Citation
Sapkota, K.,
Huang, F.
(2018). Efficient One-Pot Enzymatic Synthesis of Dephospho Coenzyme A. Bioorganic Chemistry, 76, 23-27.
Available at: https://aquila.usm.edu/fac_pubs/14919