A two-step, one-pot enzymatic synthesis of cephalexin from D-phenylglycine nitrile

• Published in: Biotechnology and bioengineering Vol. 79; no. 3; pp. 356 - 361
• Main Authors: Wegman, Margreth A., van Langen, Luuk M., van Rantwijk, Fred, Sheldon, Roger A.
• Format: Journal Article
• Language: English
• Published: New York Wiley Subscription Services, Inc., A Wiley Company 05-08-2002; Wiley
• Subjects: Acetonitriles - chemistry; Antibiotics; Biological and medical sciences; Biotechnology; Catalysis; cephalexin; Cephalexin - chemical synthesis; Cephalexin - isolation & purification; Cephalosporins - chemistry; Escherichia coli - enzymology; Fundamental and applied biological sciences. Psychology; Glycine - analogs & derivatives; Glycine - chemistry; Health. Pharmaceutical industry; Hydro-Lyases - chemistry; Industrial applications and implications. Economical aspects; Models, Chemical; Multienzyme Complexes; multienzyme synthesis; nitrile hydratase; Penicillin Amidase - chemistry; penicillin G acylase; Production of active biomolecules; Quality Control; Rhodococcus - enzymology; selective inhibition; Sensitivity and Specificity; Nitrile; Cefalexin; Phenylalanine; Enzyme; β-Lactams; Lyases; Biosynthesis; Glycine; Optimization; Penicillin amidase; Antibiotic; Cephalosporin derivatives; Biotransformation; Nitrile hydratase; Hydrolases; Antibacterial agent; Carbon-oxygen lyases; Hydro-lyases
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.10278

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. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 356–361, 2002.