A two-step, one-pot enzymatic synthesis of cephalexin from D-phenylglycine nitrile.
A cascade of two enzymatic transformations is employed in a one-pot synthesis of cephalexin. The nitrile hydratase (from R. rhodochrous MAWE)-catalyzed hydration of D-phenylglycine nitrile to the corresponding amide was combined with the penicillin G acylase (penicillin amidohydrolase, E.C. 3.5.1.11)-catalyzed acylation of 7-ADCA with the in situ-formed amide to afford a two-step, one-pot synthesis of cephalexin. D-Phenylglycine nitrile appeared to have a remarkable selective inhibitory effect on the penicillin G acylase, resulting in a threefold increase in the synthesis/hydrolysis (S/H) ratio. 1,5-Dihydroxynaphthalene, when added to the reaction mixture, cocrystallized with cephalexin. The resulting low cephalexin concentration prevented its chemical as well as enzymatic degradation; cephalexin was obtained at 79% yield with an S/H ratio of 7.7.
Wegman
Margreth A
MA
Laboratory of Biocatalysis and Organic Chemistry, Delft University of Technology, Julianalaan 136, The Netherlands.
van Langen
Luuk M
LM
van Rantwijk
Fred
F
Sheldon
Roger A
RA
D000097:Acetonitriles / Q000737:chemistry*; D002384:Catalysis; D002506:Cephalexin / Q000138:chemical synthesis* / Q000302:isolation & purification; D002511:Cephalosporins / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology; D005998:Glycine / Q000031:analogs & derivatives / Q000737:chemistry*; D006836:Hydro-Lyases / Q000737:chemistry*; D008956:Models, Chemical; D009097:Multienzyme Complexes; D010399:Penicillin Amidase / Q000737:chemistry*; D011786:Quality Control; D012240:Rhodococcus / Q000201:enzymology; D012680:Sensitivity and Specificity
D000097:Acetonitriles; D002511:Cephalosporins; D009097:Multienzyme Complexes; C470025:phenylglycinamide; C470316:phenylglycine nitrile; C014320:7-aminodesacetoxycephalosporanic acid; D010399:Penicillin Amidase; D006836:Hydro-Lyases; C048831:nitrile hydratase; D002506:Cephalexin; D005998:Glycine