Amidase from Klebsiella oxytoca

The hydrolysis with a (R)-specific amidase from Klebsiella oxytoca resolved 2-hydroxy- and 2-amino-3,3,3-trifluoro-2-methylpropionamide, giving the (R)-acid and the remaining (S)-amide (Figure 6.39) [104,105]. 

Shaw, N.M., Naughton, A., Robins, K., et al. (2002) Selection, purification, characterisation, and cloning of a novel heat-stable stereo-specific amidase from Klebsiella oxytoca, and its application in the synthesis of enantiomerically pure (R)- and (S)-3,3,3-trifluoro-2-hydroxy-2-methylpropionic acids and (S)-3,3,3-trifluoro-2-hydroxy-2-methylpropionamide. Organic Process Research Development, 6, 497-504. 

For the successful route an enantiospecific amidase from Klebsiella oxytoca was isolated, characterized, and cloned and used to resolve (R,S)-3,3,3-trifluoro-2-hy-... 

The amidase from Klebsiella oxytoca is robust, stable, and does not require cofactors. It can be heated to 70 °C for 10 min without loss of activity, and therefore, heat treatment of the biomass used in the biotransformation was used to stabilize the enzyme, presumably by inactivating proteases. 

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