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A two‐step, one‐pot enzymatic synthesis of cephalexin from D ‐phenylglycine nitrile
Author(s) -
Wegman Margreth A.,
van Langen Luuk M.,
van Rantwijk Fred,
Sheldon Roger A.
Publication year - 2002
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10278
Subject(s) - nitrile , nitrile hydratase , chemistry , rhodococcus rhodochrous , amide , hydrolysis , amidase , enzyme , penicillin , acylation , penicillin amidase , stereochemistry , yield (engineering) , nitrilase , catalysis , enzymatic hydrolysis , organic chemistry , combinatorial chemistry , biochemistry , immobilized enzyme , antibiotics , rhodococcus , materials science , metallurgy
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.