One High-Throughput Screening Strategy for Nitrile-Hydrolyzing Enzymes

The production of carboxylic acids and their derivatives which are hydrolyzed from nitrile by nitrile-hydrolyzing enzyme(nitrilase or nitrile hydratase/amidase) have been widely used in pharmaceutical.In order to obtain more nitrile-hydrolyzing enzymes(nitrilase or nitrile hydratase/amidase),it is very important to build a high-throughput screening strategy.In this paper,a modified ferric hydroxamate spectrophotometry was established,which is simple,rapid,and high-throughput.The accuracy of the method established was validated by the bioconversion of nitrile hydrolysis with the resting cells of Rhodococcus sp.CCZU10-1.The results indicate that the accuracy of this method for assaying carboxylic acids is as high as the HPLC-based method.Therefore,the method established can be used for the screening and application of microorganisms with nitrile-hydrolyzing activity.