One-Pot Multienzymatic Synthesis of 12-Ketoursodeoxycholic Acid: Subtle Cofactor Specificities Rule the Reaction Equilibria of Five Biocatalysts Working in a Row
Article first published online: 17 MAR 2009
Copyright © 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Advanced Synthesis & Catalysis
Volume 351, Issue 9, pages 1303–1311, June 2009
How to Cite
Monti, D., Ferrandi, E. E., Zanellato, I., Hua, L., Polentini, F., Carrea, G. and Riva, S. (2009), One-Pot Multienzymatic Synthesis of 12-Ketoursodeoxycholic Acid: Subtle Cofactor Specificities Rule the Reaction Equilibria of Five Biocatalysts Working in a Row. Adv. Synth. Catal., 351: 1303–1311. doi: 10.1002/adsc.200800727
- Issue published online: 16 JUN 2009
- Article first published online: 17 MAR 2009
- Manuscript Received: 21 NOV 2008
- COST Action CM0701 “Cascade Chemoenzymatic Processes - New Synergies Between Chemistry and Biochemistry”
- Prodotti Chimici Alimentari S.p.A.
- bile acids;
- hydroxysteroid dehydrogenases;
- one-pot reaction;
The hydroxysteroid dehydrogenases (HSDHs)-catalyzed one-pot enzymatic synthesis of 12-ketoursodeoxycholic acid (3α,7β-dihydroxy-12-oxo-5β-cholanoic acid), a key intermediate for the synthesis of ursodeoxycholic acid, from cholic acid has been investigated. This goal has been achieved by alternating oxidative and reductive steps in a one-pot system employing HSDHs with different cofactor specificity, namely NADH-dependent HSDHs in the oxidative step and an NADPH-dependent 7β-HSDH in the reductive one. Coupled in situ regeneration systems have been exploited not only to allow the use of catalytic amounts of the cofactors, but also to provide the necessary driving force to opposite reactions (i.e., oxidation and reduction) acting on different sites of the substrate molecule. Biocatalysts suitable for the set-up of this process have been selected and their kinetic behaviour in respect of the reactions of interest has been evaluated. Finally, the process has been studied employing the enzymes both in free and compartmentalized form.