One-pot chemoenzymatic preparation of coenzyme A analogues.
A rapid and stoichiometric method for the synthesis of analogues of coenzyme A is described. The method links the enzymes pantothenate kinase, phosphopantotheine adenylyltransferase, and dephosphocoenzyme A kinase in vitro to generate a variety of CoA analogues from chemically synthesized pantothenic acid derivatives. The Escherichia coli CoA biosynthetic enzymes were overexpressed as hexa-histidine-tagged proteins, providing an abundant source of pure active catalysts for the reaction. The synthesis of five novel CoA derivatives is reported and the method is shown to be robust and tolerant of a number of different pantothenic acid structures, which indicates that the procedure should be widely applicable.
Nazi
Ishac
I
Antimicrobial Research Centre, Department of Biochemistry, McMaster University, Ontario, Canada L8N 3Z5.
Koteva
Kalinka P
KP
Wright
Gerard D
GD
D001483:Base Sequence; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D003065:Coenzyme A / Q000096:biosynthesis* / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000378:metabolism; D010205:Pantothenic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry
D011994:Recombinant Proteins; D010205:Pantothenic Acid; C015787:dephosphocoenzyme A; D017853:Phosphotransferases (Alcohol Group Acceptor); C023088:pantothenate kinase; D009713:Nucleotidyltransferases; C022315:pantetheine-phosphate adenylyltransferase; D003065:Coenzyme A