Phosphomannose isomerase/GDP-mannose pyrophosphorylase from Pyrococcus furiosus: a thermostable biocatalyst for the synthesis of guanidinediphosphate-activated and mannose-containing sugar nucleotides.
Herein we present an analysis of the chemical function of a recombinant bifunctional phosphomannose isomerase/GDP-mannose pyrophosphorylase (manC) from Pyrococcus furiosus DSM 3638 and its use in the synthesis of guanidinediphospho-hexoses and a range of nucleotidediphospho-mannoses. This enzyme is unusually promiscuous in both its nucleotide triphosphate (NTP) and sugar-1-phosphate acceptance. It accepts all five naturally occurring NTPs (ATP, CTP, GTP, dTTP and UTP) and a range of sugar-1-phosphates (glucose-, mannose-, galactose-, glucosamine-, N-acetylglucosamine- and fucose-1-phosphate). A truncated GDP-mannose pyrophosphorylase domain of the whole length enzyme showed almost 100-fold less sugar nucleotidyltransferase activity with only GTP and mannose 1-phosphate as substrates. The temperature stability and inherently broad substrate tolerance of this archaeal enzyme make it an effective reagent for the rapid chemoenzymatic synthesis of a range of natural and unnatural sugar nucleotides that are challenging to make by chemical means alone.
Mizanur
Rahman M
RM
Department of Chemistry and Plant Sciences Institute, Gilman Hall, Iowa State University, Ames, Iowa 50011-3111, USA.
Pohl
Nicola L B
NL
D019843:Archaeal Proteins; D004795:Enzyme Stability; D004798:Enzymes / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000737:chemistry*; D008358:Mannose / Q000737:chemistry*; D008359:Mannose-6-Phosphate Isomerase / Q000302:isolation & purification / Q000378:metabolism*; D008360:Mannosephosphates; D009711:Nucleotides / Q000096:biosynthesis; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D020027:Pyrococcus furiosus / Q000201:enzymology*; D013403:Sugar Phosphates / Q000737:chemistry; D014537:Uridine Diphosphate N-Acetylglucosamine
D019843:Archaeal Proteins; D004798:Enzymes; D008360:Mannosephosphates; D009711:Nucleotides; D013403:Sugar Phosphates; D006153:Guanosine Diphosphate; C047217:mannose 1-phosphate; D014537:Uridine Diphosphate N-Acetylglucosamine; D009713:Nucleotidyltransferases; C026917:mannose 1-phosphate guanylyltransferase; D008359:Mannose-6-Phosphate Isomerase; D008358:Mannose